Spidroin Gene Research Articles

Three Novel Spider Genomes Unveil Spidroin Diversification and Hox Cluster Architecture: Ryuthela nishihirai (Liphistiidae), Uloborus plumipes (Uloboridae) and Cheiracanthium punctorium (Cheiracanthiidae).

Spiders are a hyperdiverse taxon and among the most abundant predators in nearly all terrestrial habitats. Their success is often attributed to key developments in their evolution such as silk and venom production and major apomorphies such as a whole-genome duplication. Resolving deep relationships within the spider tree of life has been historically challenging, making it difficult to measure the relative importance of these novelties for spider evolution. Whole-genome data offer an essential resource in these efforts, but also for functional genomic studies. Here, we present de novo assemblies for three spider species: Ryuthela nishihirai (Liphistiidae), a representative of the ancient Mesothelae, the suborder that is sister to all other extant spiders; Uloborus plumipes (Uloboridae), a cribellate orbweaver whose phylogenetic placement is especially challenging; and Cheiracanthium punctorium (Cheiracanthiidae), which represents only the second family to be sequenced in the hyperdiverse Dionycha clade. These genomes fill critical gaps in the spider tree of life. Using these novel genomes along with 25 previously published ones, we examine the evolutionary history of spidroin gene and structural hox cluster diversity. Our assemblies provide critical genomic resources to facilitate deeper investigations into spider evolution. The near chromosome-level genome of the 'living fossil' R. nishihirai represents an especially important step forward, offering new insights into the origins of spider traits.

Rapid production of chimeric silkworm/spider silk with improved mechanical properties by infection of nonpermissive Bombyx mori with recombinant AcMNPV harboring native-size of spidroin genes

Spider silks with excellent mechanical properties attract more attention from scientists worldwide, and the dragline silk that serves as the framework of the spider's web is considered one of the strongest fibers. However, it is unfeasible for large-scale production of spider silk due to its highly territorial, cannibalistic, predatory, and solitary behavior. Herein, to alleviate some of these problems and explore aneasy way to produce spider fibers, we constructed recombinant baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) simultaneously expressing Trichonephila clavipes native ampullate spidroin 2 (MaSp-G) and spidroin 1 (MaSp-C) driven by the promoters of silkworm fibroin genes, to infect the nonpermissive Bombyx mori larvae at the fifth instar. MaSp-G and MaSp-C were co-expressed in the posterior silk glands (PSGs) of infected silkworms and successfully secreted into the lumen of the silk gland for fibroin globule assembly. The integration of MaSp-G and MaSp-C into silkworm silk fibers significantly improved the mechanical properties of these chimeric silk fibers, especially the strength and extensibility, which may be caused by the increment of β-sheet in the chimeric silkworm/spider silk fiber. These results demonstrated that silkworms could be developed as the nonpermissive heterologous host for the mass production of chimeric silkworm/spider silk fibers via the recombinant baculovirus AcMNPV.

Transcriptome Changes Reveal the Toxic Mechanism of Cadmium and Lead Combined Exposure on Silk Production and Web-Weaving Behavior of Spider A. ventricosus.

The combined exposure of multiple metals imposes a substantial burden on the ecophysiological functions in organisms; however, the precise mechanism(s) remains largely unknown. Here, adult female A. ventricosus were exposed to single and combined exposure to cadmium (Cd) and lead (Pb) through the food chain. The aim was to explore the combined toxicity of these metals on silk production and web-weaving behavior at physiological, cellular morphological, and transcriptomic levels. The Cd and Pb combined exposure significantly inhibited the ability of silk production and web-weaving, including reduced silk fiber weight and diameter of single strands, lowered weaving position, induced nocturnal weaving, and increased instances of no-web, and showed a dose-response relationship on the Cd and Pb bioaccumulation. Concurrently, severe oxidative stress and degenerative changes in cells were observed. In addition, the combined pollution of Cd and Pb demonstrated synergistic effects, influenced by variations in concentration, on the enrichment of metals, inhibition of silk weight, oxidative damage, and cellular degeneration. At the transcriptome level, the upregulated ampullate spidroin genes and downregulated amino acid anabolic genes, upregulated Far genes and downregulated cytoskeleton-related TUBA genes, and overexpressed AChE and Glu genes may tend to present promising potential as biomarkers for silk protein synthesis, cellular degeneration, and neurotransmitter induction. This study offers an enormous capability for a comprehensive understanding of the eco-toxicological effects and mechanisms of multiheavy metals pollution.

Electrospun mini-MiSp spidroin/poly(L-lactide-co-ε-caprolactone) nanofibrous scaffolds for ARPE-19 cells

Age-related macular degeneration, a type of retinal degenerative disease, is the primary cause of severe visual impairment and irreversible blindness in people older than 50, due to age-related deterioration of the photoreceptors and the retinal pigment epithelium (RPE), which could be restored by cell therapy. However, the success of cell therapy depends on biocompatible scaffolds, which serve as carriers for the growth of RPE cells for repairing damaged or diseased retinas. In this study, the plasmid pET-SUMO-R2C for expressing a mini-spidroin, fusing the second repetitive region to the C-terminal domain of the minor ampullate spidroin gene from the spider Araneus ventricosus, was constructed and named R2C. The spidroin R2C was expressed, purified and wet spun into silk-like fibers, possessing a Young’s modulus of 3.51 ± 0.92 GPa and toughness of 24.67 ± 9.51 MPa. The spidroin R2C, mixed with poly(L-lactide-co-ε-caprolactone) (PLCL) at a ratio of 3:1, was electrospun into nanofibrous scaffolds, composed of nanometric fibers with a diameter of 751.70 ± 19.48 nm. After co-culturing with the R2C/PLCL scaffolds, the transcriptomic differences of the ARPE-19 cells were analyzed, showing that fourteen genes that are mainly related to inflammation, tumors and keratinization were upregulated more than three times, while seven genes that are mostly related to substance transport and homeostasis were downregulated more than three times. The R2C/PLCL scaffolds have the potential to be further studied to serve as a suitable carrier for RPE transplantation in the future.

A molecular atlas reveals the tri-sectional spinning mechanism of spider dragline silk

The process of natural silk production in the spider major ampullate (Ma) gland endows dragline silk with extraordinary mechanical properties and the potential for biomimetic applications. However, the precise genetic roles of the Ma gland during this process remain unknown. Here, we performed a systematic molecular atlas of dragline silk production through a high-quality genome assembly for the golden orb-weaving spider Trichonephila clavata and a multiomics approach to defining the Ma gland tri-sectional architecture: Tail, Sac, and Duct. We uncovered a hierarchical biosynthesis of spidroins, organic acids, lipids, and chitin in the sectionalized Ma gland dedicated to fine silk constitution. The ordered secretion of spidroins was achieved by the synergetic regulation of epigenetic and ceRNA signatures for genomic group-distributed spidroin genes. Single-cellular and spatial RNA profiling identified ten cell types with partitioned functional division determining the tri-sectional organization of the Ma gland. Convergence analysis and genetic manipulation further validated that this tri-sectional architecture of the silk gland was analogous across Arthropoda and inextricably linked with silk formation. Collectively, our study provides multidimensional data that significantly expand the knowledge of spider dragline silk generation and ultimately benefit innovation in spider-inspired fibers.

Chromosome-level genome and the identification of sex chromosomes in Uloborus diversus.

The orb web is a remarkable example of animal architecture that is observed in families of spiders that diverged over 200 million years ago. While several genomes exist for araneid orb-weavers, none exist for other orb-weaving families, hampering efforts to investigate the genetic basis of this complex behavior. Here we present a chromosome-level genome assembly for the cribellate orb-weaving spider Uloborus diversus. The assembly reinforces evidence of an ancient arachnid genome duplication and identifies complete open reading frames for every class of spidroin gene, which encode the proteins that are the key structural components of spider silks. We identified the 2 X chromosomes for U. diversus and identify candidate sex-determining loci. This chromosome-level assembly will be a valuable resource for evolutionary research into the origins of orb-weaving, spidroin evolution, chromosomal rearrangement, and chromosomal sex determination in spiders.

Classification and functional characterization of spidroin genes in a wandering spider, Pardosa pseudoannulata

Spiders impress us with their sophisticated use of silk and the stunningly distinct silk proteins (spidroins) in each spider species. Understanding how silks and spidroins function and evolve within the spider world is one profound interest to expand our knowledge on spider evolution. Spidroins are characterized with the divergent repeat core region flanked with the relatively conserved N- and C-terminus. The structure and number of the repeats contribute to the unique mechanical properties of the spidroin and the silk. Spidroins have been intensively studied in web-weaver spiders, but information regarding their diversity in wandering spiders remains scarce. Here, twenty spidroin genes were identified in the pond wolf spider, Pardosa pseudoannulata, belonging to the retrolateral tibial apophysis (RTA) clade. These spidroins were categorized into four classes, including twelve ampullate spidroin (AmpSp), four aciniform spidroin (AcSp), one tubuliform spidroin (TuSp), one pyriform spidroin (PiSp), and two spidroin-like proteins. Multiple copies of the AmpSp and AcSp genes were tandemly arranged in a cluster within the genome, and the N-terminal domains and repetitive sequences of the proximately located spidroins were highly similar, suggesting that the spidroin genes diversified via tandem duplication. Only four types of morphologically distinct silk glands were found in P. pseudoannulata, namely Ma, Mi, Ac, and Pi glands, consistent with the glandular affiliation hypothesis that spidroins co-evolved with glandular specialization to fit species-specific needs. Expression profiling revealed that the single tubuliform spidroin (TuSp) gene was highly expressed in gravid females and two AcSp genes displayed synchronous expression. Knock-down of the TuSp gene via RNAi resulted in fragile and cracked eggsacs and prolonged the female pre-oviposition period, validating its importance in spider reproduction. The genome-scale characterization and functional study of spidroin genes allows associating the presence of specific spidroins with silk utility in P. pseudoannulata and will expand our knowledge of spider evolution.

Characterization of the genome and silk-gland transcriptomes of Darwin's bark spider (Caerostris darwini).

Natural silks crafted by spiders comprise some of the most versatile materials known. Artificial silks-based on the sequences of their natural brethren-replicate some desirable biophysical properties and are increasingly utilized in commercial and medical applications today. To characterize the repertoire of protein sequences giving silks their biophysical properties and to determine the set of expressed genes across each unique silk gland contributing to the formation of natural silks, we report here draft genomic and transcriptomic assemblies of Darwin's bark spider, Caerostris darwini, an orb-weaving spider whose dragline is one of the toughest known biomaterials on Earth. We identify at least 31 putative spidroin genes, with expansion of multiple spidroin gene classes relative to the golden orb-weaver, Trichonephila clavipes. We observed substantial sharing of spidroin repetitive sequence motifs between species as well as new motifs unique to C. darwini. Comparative gene expression analyses across six silk gland isolates in females plus a composite isolate of all silk glands in males demonstrated gland and sex-specific expression of spidroins, facilitating putative assignment of novel spidroin genes to classes. Broad expression of spidroins across silk gland types suggests that silks emanating from a given gland represent composite materials to a greater extent than previously appreciated. We hypothesize that the extraordinary toughness of C. darwini major ampullate dragline silk may relate to the unique protein composition of major ampullate spidroins, combined with the relatively high expression of stretchy flagelliform spidroins whose union into a single fiber may be aided by novel motifs and cassettes that act as molecule-binding helices. Our assemblies extend the catalog of sequences and sets of expressed genes that confer the unique biophysical properties observed in natural silks.

Characterization of two full-length Araneus ventricosus major ampullate silk protein genes

Major ampullate silk is noted for its great tensile strength and extensibility. The impressive material properties of major ampullate silk result from their component proteins that encoded by members of the spidroin (spider fibroin) gene family. Although the major ampullate spidroin type has evolved multiple variants within specific-species, most sequences are fragmented. Here, we present two complete major ampullate spidroin genes from the orb-weaving spider Araneus ventricosus. Due to the abundant GPG motifs in their repetitive region, the two MaSp genes were grouped in MaSp2 subclass and named MaSp2C and MaSp2D, respectively. Analysis of the full-length gene sequences reveals that both of them include a single enormous exon (10,851 bp for MaSp2C and 8640 bp for MaSp2D) that mainly translates into a central repetitive region containing multiple amino acid motifs that can be organized into five ensemble types. We use gene-specific PCR primers to search the cDNA from major ampullate glands and find evidence for alternative splicing of MaSp2D transcripts into a minor spliceoform lacking the entire repetitive domain as well as the partial terminal regions. Our results not only provide new templates for protein-based materials with tailored properties, but suggest gene and transcriptional diversity of major ampullate silk.

Chromosome-level genome assembly of the black widow spider Latrodectus elegans illuminates composition and evolution of venom and silk proteins.

BackgroundThe black widow spider has both extraordinarily neurotoxic venom and three-dimensional cobwebs composed of diverse types of silk. However, a high-quality reference genome for the black widow spider was still unavailable, which hindered deep understanding and application of the valuable biomass.FindingsWe assembled the Latrodectus elegans genome, including a genome size of 1.57 Gb with contig N50 of 4.34 Mb and scaffold N50 of 114.31 Mb. Hi-C scaffolding assigned 98.08% of the genome to 14 pseudo-chromosomes, and with BUSCO, completeness analysis revealed that 98.4% of the core eukaryotic genes were completely present in this genome. Annotation of this genome identified that repetitive sequences account for 506.09 Mb (32.30%) and 20,167 protein-coding genes, and specifically, we identified 55 toxin genes and 26 spidroins and provide preliminary analysis of their composition and evolution.ConclusionsWe present the first chromosome-level genome assembly of a black widow spider and provide substantial toxin and spidroin gene resources. These high-qualified genomic data add valuable resources from a representative spider group and contribute to deep exploration of spider genome evolution, especially in terms of the important issues on the diversification of venom and web-weaving pattern. The sequence data are also firsthand templates for further application of the spider biomass.

Expression of spidroin proteins in the silk glands of golden orb-weaver spiders.

The expression of spidroins in the major ampullate, minor ampullate, flagelliform, and tubuliform silk glands of Trichonephila clavipes spiders was analyzed using proteomics analysis techniques. Spidroin peptides were identified and assigned to different gene products based on sequence concurrence when compared with the whole genome of the spider. It was found that only a relatively low proportion of the spidroin genes are expressed as proteins in any of the studied glands. In addition, the expression of spidroin genes in different glands presents a wide range of patterns, with some spidroins being found in a single gland exclusively, while others appear in the content of several glands. The combination of precise genomics, proteomics, microstructural, and mechanical data provides new insights both on the design principles of these materials and how these principles might be translated for the production of high-performance bioinspired artificial fibers.

Gene expression profiling reveals candidate genes for defining spider silk gland types

Molecular studies of the secretory glands involved in spider silk production have revealed candidate genes for silk synthesis and a complicated history of spider silk gene evolution. However, differential gene expression profiles of the multiple silk gland types within an individual orb-web weaving spider are lacking. Each of these gland types produces a functionally distinct silk type. Comparison of gene expression among spider silk gland types would provide insight into the genes that define silk glands generally from non-silk gland tissues, and the genes that define silk glands from each other. Here, we perform 3’ tag digital gene expression profiling of the seven silk gland types of the silver garden orb weaver Argiope argentata. Five of these gland types produce silks that are non-adhesive fibers, one silk includes both fibers and glue-like adhesives, and one silk is exclusively glue-like. We identify 1275 highly expressed, significantly upregulated, and tissue specific silk gland specific transcripts (SSTs). These SSTs include seven types of spider silk protein encoding genes known as spidroin genes. We find that the fiber-producing major ampullate and minor ampullate silk glands have more similar expression profiles than any other pair of glands. We also find that a subset of the SSTs is enriched for transmembrane transport and oxidoreductases, and that these transcripts highlight differences and similarities among the major ampullate, minor ampullate, and aggregate silk glands. Furthermore, we show that the wet glue-producing aggregate glands have the most unique SSTs, but still share some SSTs with fiber producing glands. Aciniform glands were the only gland type to share a majority of SSTs with other silk gland types, supporting previous hypotheses that duplication of aciniform glands and subsequent divergence of the duplicates gave rise to the multiple silk gland types within an individual spider.

Repeat Variation Resolves a Complete Aggregate Silk Sequence of Bolas Spider Mastophora phrynosoma.

Many species of spider use a modified silk adhesive, called aggregate glue, to aid in prey capture. Aggregate spidroins (spider fibroins) are modified members of the spider silk family; however, they are not spun into fibers as are their solid silk relatives. The genes that encode for aggregate spidroins are the largest of the known spidroin genes and are similarly highly repetitive. In this study, we used long read sequencing to discover the aggregate spidroin genes of the toad-like bolas spider, Mastophora phrynosoma, which employs the glue in a unique way, using only a single, large droplet to capture moths. While Aggregate Spidroin 1 (AgSp1) remains incomplete, AgSp2 is more than an extraordinary 62 kb of coding sequence, 20 kb longer than the longest spidroin on record. The structure of repeats from both aggregate silk proteins follows a similar pattern seen in other species, with the same strict conservation of amino acid residue number for much of the repeats' lengths. Interestingly, AgSp2 lacks the elevated number and groupings of glutamine residues seen in the other reported AgSp2 of a classic orb weaving species. The role of gene length in glue functionality remains a mystery, and thus discovering length differences across species will allow understanding and harnessing of this attribute for the next generation of bio-inspired adhesives.

Transgenic Cotton Plants Expressing Spidroin Gene Presenting Increased Fiber Quality

Introduction: Spider silks are the strongest and most elastic fibers known in the nature. Cotton is also one of the most important crops in the world which increasing the quality of its fibers is of great importance in the textile industry, so the production of longer and stronger cotton fibers is of great importance. The aim of this study was to produce transgenic cotton fibers containing MaSp1 cobweb protein. Material and Methods: A synthetic construct was designed based on a selected fragment of Latrodectushesperus MaSp1 gene. The gene was then cloned twice inpCAMBIA1304 binary vector using different restriction enzymes (NcoI and NheI; EcoR1 and NheI). Agrobacterium tumefaciens LBA4404strain was transformed by recombinant vectors which are used to transform cotton plants. The accuracy of the results was confirmed using PCR, hygromycin resistance gene and recombinant protein expression. Finally, the fiber strength of transgenic cotton was measured by two vectors. Result: MaSp1 synthetic gene was transferred to the plant meristem. The study of transgenic plants in the next generations confirmed the accuracy of gene transfer and expression of recombinant protein. Conclusion: Considering the economic role of cotton in the world and also having the third rank among the industrial plants of the country, this study succeeded in producing transgenic cotton plants expressing Masp1 protein with increased fiber quality.

Golden orb-weaving spider (Trichonephila clavipes) silk genes with sex-biased expression and atypical architectures.

Spider silks are renowned for their high-performance mechanical properties. Contributing to these properties are proteins encoded by the spidroin (spider fibroin) gene family. Spidroins have been discovered mostly through cDNA studies of females based on the presence of conserved terminal regions and a repetitive central region. Recently, genome sequencing of the golden orb-web weaver, Trichonephila clavipes, provided a complete picture of spidroin diversity. Here, we refine the annotation of T. clavipes spidroin genes including the reclassification of some as non-spidroins. We rename these non-spidroins as spidroin-like (SpL) genes because they have repetitive sequences and amino acid compositions like spidroins, but entirely lack the archetypal terminal domains of spidroins. Insight into the function of these spidroin and SpL genes was then examined through tissue- and sex-specific gene expression studies. Using qPCR, we show that some silk genes are upregulated in male silk glands compared to females, despite males producing less silk in general. We also find that an enigmatic spidroin that lacks a spidroin C-terminal domain is highly expressed in silk glands, suggesting that spidroins could assemble into fibers without a canonical terminal region. Further, we show that two SpL genes are expressed in silk glands, with one gene highly evolutionarily conserved across species, providing evidence that particular SpL genes are important to silk production. Together, these findings challenge long-standing paradigms regarding the evolutionary and functional significance of the proteins and conserved motifs essential for producing spider silks.

The common house spider, Parasteatoda tepidariorum, maintains silk gene expression on sub-optimal diet.

Cobweb weaving spiders and their relatives spin multiple task-specific fiber types. The unique material properties of each silk type result from differences in amino acid sequence and structure of their component proteins, primarily spidroins (spider fibrous proteins). Amino acid content and gene expression measurements of spider silks suggest some spiders change expression patterns of individual protein components in response to environmental cues. We quantified mRNA abundance of three spidroin encoding genes involved in prey capture in the common house spider, Parasteatoda tepidariorum (Theridiidae), fed different diets. After 10 days of acclimation to the lab on a diet of mealworms, spiders were split into three groups: (1) individuals were immediately dissected, (2) spiders were fed high-energy crickets, or (3) spiders were fed low-energy flies, for 1 month. All spiders gained mass during the acclimation period and cricket-fed spiders continued to gain mass, while fly-fed spiders either maintained or lost mass. Using quantitative PCR, we found no significant differences in the absolute or relative abundance of dragline gene transcripts, major ampullate spidroin 1 (MaSp1) and major ampullate spidroin 2 (MaSp2), among groups. In contrast, prey-wrapping minor ampullate spidroin (MiSp) gene transcripts were significantly less abundant in fly-fed than lab-acclimated spiders. However, when measured relative to Actin, cricket-fed spiders showed the lowest expression of MiSp. Our results suggest that house spiders are able to maintain silk production, even in the face of a low-quality diet.

Long-read transcriptomic analysis of orb-weaving spider Araneus ventricosus indicates transcriptional diversity of spidroins

Spider silk, which is composed of diverse silk proteins (spidroin), is a kind of natural high-mass biomaterial with great potential. However, due to the complexity of both the structure and the composition of the spidroins in natural spider silk, application of this valuable biomass is still limited to date. There are diverse kinds of spider silk in the orb-weaving spider with different mechanical and structural characteristics. In order to systematically illustrate the landscape of all the different spidrons, here we chose Araneus ventricosus, an orb-weaving spider with superior silk mechanical features and genome information, to generate a long-read whole body transcriptome. We deciphered the repeat arrangements of each kind of spidroin, based on which we found that there are substantially transcriptional diversity of each spidroin gene. Some repeat motifs are not documented before. Specifically, we discovered novel full-lengh MaSp transcript as well as a relatively small full-length AcSp isoforms, which are potential promising materials for bioengineering of recombinant spidroin. Our study provided a batch of new spidron resources with detail sequential information. The finding of transcriptional diversity may provide cues in understanding of within-species variation of the mechanical properties of the natural spider silk and further molecular designing of recombinant spidroin.

Spidroin profiling of cribellate spiders provides insight into the evolution of spider prey capture strategies

Orb-weaving spiders have two main methods of prey capture: cribellate spiders use dry, sticky capture threads, and ecribellate spiders use viscid glue droplets. Predation behaviour is a major evolutionary driving force, and it is important on spider phylogeny whether the cribellate and ecribellate spiders each evolved the orb architecture independently or both strategies were derived from an ancient orb web. These hypotheses have been discussed based on behavioural and morphological characteristics, with little discussion on this subject from the perspective of molecular materials of orb web, since there is little information about cribellate spider-associated spidroin genes. Here, we present in detail a spidroin catalogue of six uloborid species of cribellate orb-weaving spiders, including cribellate and pseudoflagelliform spidroins, with transcriptome assembly complemented with long read sequencing, where silk composition is confirmed by proteomics. Comparative analysis across families (Araneidae and Uloboridae) shows that the gene architecture, repetitive domains, and amino acid frequencies of the orb web constituting silk proteins are similar among orb-weaving spiders regardless of the prey capture strategy. Notably, the fact that there is a difference only in the prey capture thread proteins strongly supports the monophyletic origin of the orb web.

Spider Silks: An Overview of Their Component Proteins for Hydrophobicity and Biomedical Applications.

Spider silks have received extensive attention from scientists and industries around the world because of their remarkable mechanical properties, which include high tensile strength and extensibility. It is a leading-edge biomaterial resource, with a wide range of potential applications. Spider silks are composed of silk proteins, which are usually very large molecules, yet many silk proteins still remain largely underexplored. While there are numerous reviews on spider silks from diverse perspectives, here we provide a most up-to-date overview of the spider silk component protein family in terms of its molecular structure, evolution, hydrophobicity, and biomedical applications. Given the confusion regarding spidroin naming, we emphasize the need for coherent and consistent nomenclature for spidroins and provide recommendations for pre-existing spidroin names that are inconsistent with nomenclature. We then review recent advances in the components, identification, and structures of spidroin genes. We next discuss the hydrophobicity of spidroins, with particular attention on the unique aquatic spider silks. Aquatic spider silks are less known but may inspire innovation in biomaterials. Furthermore, we provide new insights into antimicrobial peptides from spider silk glands. Finally, we present possibilities for future uses of spider silks.

Characterization of the second type of aciniform spidroin (AcSp2) provides new insight into design for spidroin-based biomaterials

Spiders spin a range of silks from different glands for distinct functions, and each silk type exhibits distinct material properties. Silk extruded by the aciniform gland is used for prey wrapping and egg case construction and displays high toughness and extensibility. So far, only the aciniform spidroin 1 (AcSp1) gene which was firstly identified as a silk gene in aciniform gland has been obtained. Here we present the gene sequence for the second type of full-length aciniform silk protein, AcSp2. Analysis of the AcSp2 primary sequence reveals relatively conserved terminal regions and a distinct repetitive sequence relative to AcSp1. A fraction of the gene can be expressed in recombinant systems. Secondary structure analysis of the recombinant AcSp2 protein in solution reveals that the protein adopts mainly an α-helical conformation. Artificial spinning of recombinant AcSp2 demonstrates that the spidroins can be spun into fine fibers which display up to 142% extensibility. The silk fibers are dominated by β-sheet and β-turn secondary structures. Moreover, the mechanical data collected from these synthetic fibers revealed that the mechanical properties are partly correlated with the molecular weights. Overall, our studies enrich our knowledge of spidroin gene family members and provide a new insight into creation of high-performance silk fibers for next generation biomaterials. Statement of SignificanceIn this study, we presented the second type of aciniform silk protein (AcSp2) gene sequence of orb-weaving spider Araneus ventricosus, expanding the spider silk gene family members. The primary structure revealed the central repetitive sequence of the new spidroin gene is distinctly different from other AcSp1 genes. Characterization of the recombinant minispidroin fibers of AcSp2 revealed the mechanical properties are partly correlate with the molecular weights, and the spidroins can be spun into fine fibers which display up to 142% extensibility. Overall, our studies enrich our knowledge of spidroin gene family members and provide a new insight into creation of high-performance silk fibers for next generation biomaterials.