Antiparallel Sheet Research Articles

Assessment of structural behaviour of a new L-asparaginase and SAXS data-based evidence for catalytic activity in its monomeric form

The present study reports the structural and functional characterization of a new glutaminase-free recombinant L-asparaginase (PrASNase) from Pseudomonas resinovorans IGS-131. PrASNase showed substrate specificity to L-asparagine, and its kinetic parameters, Km, Vmax, and kcat were 9.49 × 10−3 M, 25.13 IUmL−1 min−1, and 3.01 × 103 s−1, respectively. The CD spectra showed that PrASNase consisted of 18.5 % helix, 21.5 % antiparallel sheets, 4.2 % parallel sheets, 14 % turns, and rest other structures. FTIR was used for the functional characterization, and molecular docking predicted that the substrate interacts with serine, alanine, and glutamine in the binding pocket of PrASNase. Differing from known asparaginases, structural characterization by small-angle X-ray scattering (SAXS) and analytical ultracentrifugation (AUC) unambiguously revealed PrASNase to exist as a monomer in solution at low temperatures and oligomerized to a higher state with temperature rise. Through SAXS studies and enzyme assay, PrASNase was found to be mostly monomer and catalytically active at 37 °C. Furthermore, this glutaminase-free PrASNase showed killing effects against WIL2-S and TF-1.28 cells with IC50 of 7.4 μg.mL−1 and 5.6 μg.mL−1, respectively. This is probably the first report with significant findings of fully active L-asparaginase in monomeric form using SAXS and AUC and demonstrated the potential of PrASNase in inhibiting cancerous cells, making it a potential therapeutic candidate.

A Hairpin Motif in the Amyloid-β Peptide Is Important for Formation of Disease-Related Oligomers.

The amyloid-β (Aβ) peptide is associated with the development of Alzheimer's disease and is known to form highly neurotoxic prefibrillar oligomeric aggregates, which are difficult to study due to their transient, low-abundance, and heterogeneous nature. To obtain high-resolution information about oligomer structure and dynamics as well as relative populations of assembly states, we here employ a combination of native ion mobility mass spectrometry and molecular dynamics simulations. We find that the formation of Aβ oligomers is dependent on the presence of a specific β-hairpin motif in the peptide sequence. Oligomers initially grow spherically but start to form extended linear aggregates at oligomeric states larger than those of the tetramer. The population of the extended oligomers could be notably increased by introducing an intramolecular disulfide bond, which prearranges the peptide in the hairpin conformation, thereby promoting oligomeric structures but preventing conversion into mature fibrils. Conversely, truncating one of the β-strand-forming segments of Aβ decreased the hairpin propensity of the peptide and thus decreased the oligomer population, removed the formation of extended oligomers entirely, and decreased the aggregation propensity of the peptide. We thus propose that the observed extended oligomer state is related to the formation of an antiparallel sheet state, which then nucleates into the amyloid state. These studies provide increased mechanistic understanding of the earliest steps in Aβ aggregation and suggest that inhibition of Aβ folding into the hairpin conformation could be a viable strategy for reducing the amount of toxic oligomers.

A novel diG motif in ORF3a protein of SARS-Cov-2 for intracellular transport.

The ongoing SARS-CoV-2/COVID-19 pandemic caused a global public health crisis. Yet, everyone's response to SARS-CoV-2 infection varies, and different viral variants confer diverse pathogenicity. Thus, it is imperative to understand how viral determinants contribute to COVID-19. Viral ORF3a protein is one of those viral determinants, as its functions are linked to induction of cell and tissues damages, disease severity and cytokine storm that is a major cause of COVID-19-related death. ORF3a is a membrane-associated protein. Upon synthesis, it is transported from endoplasmic reticulum, Golgi apparatus to plasma membrane and subcellular endomembranes including endosomes and lysosomes. However, how ORF3a is transported intracellularly remains elusive. The goal of this study was to carry out a systematic mutagenesis study to determine the structural relationship of ORF3a protein with its subcellular locations. Single amino acid (aa) and deletion mutations were generated in the putative function-relevant motifs and other regions of interest. Immunofluorescence and ImageJ analyses were used to determine and quantitate subcellular locations of ORF3a mutants in comparison with wildtype ORF3a. The wildtype ORF3a localizes predominantly (Pearson's coefficients about 0.8) on the membranes of endosomes and lysosomes. Consistent with earlier findings, deletion of the YXXΦ motif, which is required for protein export, retained ORF3a in the Golgi apparatus. Interestingly, mutations in a double glycine (diG) region (aa 187-188) displayed a similar phenotype to the YXXΦ deletion, implicating a similar role of the diG motif in intracellular transport. Indeed, interrupting any one of the two glycine residues such as deletion of a single (dG188), both (dG187/dG188) or substitution (G188Y) of these residues led to ORF3a retention in the Golgi apparatus (Pearson's coefficients ≥0.8). Structural analyses further suggest that the diG motif supports a type-II β-turn between the anti-parallel β4 and β5 sheets and connects to the YXXΦ motif via hydrogen bonds between two monomers. The diG- YXXΦ interaction forms a hand-in-hand configuration that could facilitate dimerization. Together, these observations suggest a functional role of the diG motif in intracellular transport of ORF3a.

High-resolution crystal structure of the anti-CRISPR protein AcrIC5

As a result of the long-term battle of bacteria and archaea against invaders such as viruses and genetic mobile elements, they have developed CRISPR-Cas systems for self-defense, which allows them to remove the viral genetic material introduced into host cells via infection. To fight against this bacterial immune system, however, viruses have also evolved to produce multiple anti-CRISPR proteins that can inhibit the bacterial CRISPR-Cas system. In this study, we introduced a tentative inhibitory activity against a type I–C CRISPR-Cas system by determining the crystal structure of AcrIC5 from Pseudomonas delhiensis. Structural analysis revealed that AcrIC5 was composed of noble folds comprising two antiparallel sheets and three helices. Although AcrIC5 did not directly interact with either the type I–C cascade from Neisseria lactamia or the type I–F cascade from Pseudomonas aeruginosa in our analysis, a highly acidic surface feature indicated that AcrIC5 may be DNA mimic Acrs that directly binds to the target DNA binding site in type I–C cascade and inhibits the recruitment of the target DNA to this cascade.

CgIL17-5 regulates the mRNA expressions of immune effectors through inducing the phosphorylation of CgMAPKs and the nuclear translocation of CgRel and CgAP-1 in the Pacific oyster Crassostrea gigas

Interleukin-17 (IL-17) is a classic pro-inflammatory cytokine that plays an important role in the immune and inflammatory response. In the present study, the sequence feature of CgIL17-5 and its function as a pro-inflammatory factor in inducing the mRNA expressions of downstream immune effectors were investigated in oyster Crassostrea gigas. There were two tightly folded alpha helixes and two pairs of antiparallel beta-pleated sheet in the amino acid sequence of CgIL17-5. The mRNA transcripts of CgIL17-5 were constitutively distributed in all the tested tissues, with the highest level in haemocytes. The mRNA expression level of CgIL17-5 in haemocytes increased significantly at 24 h after Vibrio splendidus stimulation. CgIL17-5 protein was mainly detected in granulocytes which were the main immunocompetent haemocytes in C. gigas. The phosphorylation of mitogen-activated protein kinases (CgJNK, CgERK and CgP38) and nuclear translocation of the transcription factors (CgRel and CgAP-1) in haemocytes were induced after the oysters received an injection of recombinant CgIL17-5 for 2 h. The mRNA expression levels of CgIL-17s, CgTNF-1, Cgdefh1 and Cgdefh2 increased significantly in haemocytes. At the same time, obvious branchial swelling and cilium shedding in gills were observed at 24 h after the oysters received an injection of rCgIL17-5. All the results collectively suggested that CgIL17-5 promoted the activation of CgMAPKs and the nuclear translocation of CgRel and CgAP-1 to promote the mRNA expressions of cytokines and antibacterial peptides.

Constraints on the Structure of Fibrils Formed by a Racemic Mixture of Amyloid-β Peptides from Solid-State NMR, Electron Microscopy, and Theory.

Previous studies have shown that racemic mixtures of 40- and 42-residue amyloid-β peptides (d,l-Aβ40 and d,l-Aβ42) form amyloid fibrils with accelerated kinetics and enhanced stability relative to their homochiral counterparts (l-Aβ40 and l-Aβ42), suggesting a "chiral inactivation" approach to abrogating the neurotoxicity of Aβ oligomers (Aβ-CI). Here we report a structural study of d,l-Aβ40 fibrils, using electron microscopy, solid-state nuclear magnetic resonance (NMR), and density functional theory (DFT) calculations. Two- and three-dimensional solid-state NMR spectra indicate molecular conformations in d,l-Aβ40 fibrils that resemble those in known l-Aβ40 fibril structures. However, quantitative measurements of 13C-13C and 15N-13C distances in selectively labeled d,l-Aβ40 fibril samples indicate a qualitatively different supramolecular structure. While cross-β structures in mature l-Aβ40 fibrils are comprised of in-register, parallel β-sheets, our data indicate antiparallel β-sheets in d,l-Aβ40 fibrils, with alternation of d and l molecules along the fibril growth direction, i.e., antiparallel "rippled sheet" structures. The solid-state NMR data suggest the coexistence of d,l-Aβ40 fibril polymorphs with three different registries of intermolecular hydrogen bonds within the antiparallel rippled sheets. DFT calculations support an energetic preference for antiparallel alignments of the β-strand segments identified by solid-state NMR. These results provide insight into the structural basis for Aβ-CI and establish the importance of rippled sheets in self-assembly of full-length, naturally occurring amyloidogenic peptides.

Unconventional Secondary Structure Mimics: Ladder‐Rungs

AbstractSecondary structures tend to be recognizable because they have repeating structural motifs, but mimicry of these does not have to follow such well‐defined patterns. Bioinformatics studies to match side‐chain orientations of a novel hydantoin triazole chemotype (1) to protein‐protein interfaces revealed it tends to align well across parallel and antiparallel sheets, like rungs on a ladder. One set of these overlays was observed for the protein‐protein interaction uPA⋅uPAR. Consequently, chemotype 1 was made with appropriate side‐chains to mimic uPA at this interface. Biophysical assays indicate these compounds did in fact bind uPAR, and elicit cellular responses that affected invasion, migration, and wound healing.

Evolutionary Analysis of Pectin Lyases of the Genus Colletotrichum.

Pectin lyases (PNLs) are important enzymes that are involved in plant cell wall degradation during the infection process. Colletotrichum is a diverse genus of fungi, which allows the study of the evolution of PNLs and their possible role in pathogen-host interactions and lifestyle adaptations. The phylogenetic reconstruction of PNLs from Colletotrichum and analysis of selection pressures showed the formation of protein lineages by groups of species with different selection pressures and specific patterns. The analysis of positive selection at individual sites using different methods allowed for the identification of three codons with evidence of positive selection in the oligosaccharide-binding region and two codons on the antiparallel sheet, which may influence the interaction with the substrate. Seven codons on the surface of the protein, mainly in the peripheral helices of the PNLs, could have an important function in evasion of plant defenses, as has been proposed in other enzymes. According to our results, it is possible that events of genetic duplication occurred in ancestral lines, followed by episodes of genetic diversification and gene loss, probably influenced by differences in the composition of the host cell wall. Additionally, different patterns of evolution in Colletotrichum appear to be molded by a strong purifying selection and positive selection episodes that forged the observed evolutionary patterns, possibly influenced by host interaction or substrate specificity. This work represents a starting point for the study of sites that may be important for evasion of plant defenses and biotechnological purposes.

An Atypical Place for Amyloidosis

Amyloidosis is the abnormal extracellular tissue deposition of insoluble, pathologic fibrils composed of antiparallel beta-pleated sheets. Deposition can be systemic or organ-specific. Symptomatic gastric amyloidosis is rare given that only 0.08% of those afflicted with the disease demonstrate gastrointestinal bleeding, malabsorption, protein-losing gastroenteropathy, and chronic gastrointestinal dysmotility. A known correlation exists between multiple myeloma and amyloidosis. This is a case of a 75 year old male with multiple myeloma who was found to have gastric and duodenal amyloidosis on evaluation of nausea, abdominal pain and dysphagia with EGD. For his multiple myeloma the patient underwent treatment for several months with dexamethasone, cyclophosphamide, and bortezomide. On a follow up visit, he endorsed several weeks of nausea, epigastric abdominal pain and dysphagia (mainly to solids) and thus EGD was performed. He was found to have a mild, non-obstructing Schatzki ring at the gastroesophageal junction. It was dilated to a maximum TTS balloon size of 20mm with complete resolution of luminal narrowing. He was also found to have localized moderately erythematous mucosa without bleeding in the gastric antrum as well as two 5mm non-bleeding cratered duodenal bulb ulcers with no stigmata of bleeding. These sites were biopsied and confirmed to have abundant amyloid deposition in the lamina propria via the Congo red special stain. Amyloid deposition can occur in the lower esophageal sphincter thereby decreasing lower esophageal sphincter pressure and cause symptoms of dysphagia and acid reflux, however typical symptoms of gastrointestinal amyloidosis include hematochezia, melena, malabsorption and dysmotility. Gastric amyloidosis, though rare, should be considered on differential diagnosis in individuals with known multiple myeloma who present with nausea, abdominal pain and dysphagia.

POSS-Appended Diphenylalanine: Self-Cleaning, Pollution-Protective, and Fire-Retardant Hybrid Molecular Material.

Ordered self-assemblies of hybrid molecules have potential as protective umbrellas against environmental pollution and corrosion. This article describes the design and fabrication of a new self-cleaning hybrid molecular material containing polyhedral oligomeric silsesquioxane (POSS) and diphenylalanine as hydrophobic and pollution-protective coating. The colorless organic–inorganic hybrid materials, in which the diphenylalanine motif controls the self-assembly, exhibit unique water resistance property and enhance mechanical strength of paper 1.5-fold. The hybrid building blocks self-assemble in antiparallel sheet manner through noncovalent interactions to form a supramolecular layerlike surface with enhanced roughness. The hybrid material is soluble in organic solvents at room temperature that makes it easy to coat on paper, wood, or metal surfaces. The coating is effective against rusting, corrosion, environmental pollution, and bacterial attack. The coating has been used as fire retardant and enhances fire safety. The sustainable molecular material is promising for the packaging industry and metal industry and artifact preservation.

Conformational Effects through Hydrogen Bonding in a Constrained γ-Peptide Template: From Intraresidue Seven-Membered Rings to a Gel-Forming Sheet Structure.

A series of three short oligomers (di-, tri-, and tetramers) of cis-2-(aminomethyl)cyclobutane carboxylic acid, a γ-amino acid featuring a cyclobutane ring constraint, were prepared, and their conformational behavior was examined spectroscopically and by molecular modeling. In dilute solutions, these peptides showed a number of low-energy conformers, including ribbonlike structures pleated around a rarely observed series of intramolecular seven-membered hydrogen bonds. In more concentrated solutions, these interactions defer to an organized supramolecular assembly, leading to thermoreversible organogel formation notably for the tripeptide, which produced fibrillar xerogels. In the solid state, the dipeptide adopted a fully extended conformation featuring a one-dimensional network of intermolecularly H-bonded molecules stacked in an antiparallel sheet alignment. This work provides unique insight into the interplay between inter- and intramolecular H-bonded conformer topologies for the same peptide template.

Tautomeric Effect of Histidine on the Monomeric Structure of Amyloid β-Peptide(1-40).

Histidine state (deprotonated, neutral, and protonated) is considered an important factor influencing the structural properties and aggregation mechanisms in amyloid β-peptides (Aβ), which are associated with the pathogenesis of Alzheimer's disease. Understanding the structural properties and aggregation mechanisms is a great challenge because two forms (the Nε-H or Nδ-H tautomer) can exist in the free neutral state of histidine. Here, replica-exchange molecular dynamics simulation was performed to elucidate the changes in structure and the mechanism of aggregation influenced by tautomeric behaviors of histidine in Aβ(1-40). Our results show that sheet-dominating conformations can be found in the His6(δ)-His13(δ)-His14(δ) (δδδ) isomer with significant antiparallel sheet structures between R5-D7 and L34-G38, as well as between L17-F20 and L34-G38, implying that a new aggregation mechanism may exist to promote the generation of oligomers and/or aggregates. This work is helpful in understanding the fundamental tautomeric behaviors of neutral histidine in the process of aggregation.

Thermophysical Properties of Anti-Parallel β-Sheets with <i>Bombyx mori</i> Silk Nanostructures [Gly-Ser-Gly-Ala-Gly-Ala]<sub>n</sub> and [Gly-Ala]<sub>n</sub>

Thermal expansion of anti-parallel beta pleated sheets with Bombyx mori silk nanostructures [Gly-Ser-Gly-Ala-Gly-Ala]n and commonly used model of [Gly-Ala]n have been investigated through molecular dynamics (MD) simulation calculations in conjunction with interatomic interactions modeled by CHARMM force field method between the temperature range of 300K-700K. Preliminary results indicate significant differences on thermal expansion of two structures that was observed on directions of chain and the direction perpendicular to plates.

Thermomechanical Properties of Anti-Parallel β-Sheets with <i>Bombyx mori</i> Silk Nanostructures [Gly-Ser-Gly-Ala-Gly-Ala]<sub>n</sub> and [Gly-Ala]<sub>n</sub>

The present study aims to investigate temperature dependence of anisotropic mechanical properties of anti-parallel beta pleated sheets with Bombyx mori silk nanostructures in sequence of [Gly-Ser-Gly-Ala-Gly-Ala]n and commonly used model in sequence of [Gly-Ala]n. Classical molecular dynamics simulation method was used to carry out calculations, preliminary, at 300K. Ionic interactions were modeled by CHARMM force field method. Substitution of Ala with Ser residue results with a gradual conformational change in the stress-strain.

Role of Side Chains in β-Sheet Self-Assembly into Peptide Fibrils. IR and VCD Spectroscopic Studies of Glutamic Acid-Containing Peptides.

Poly(glutamic acid) at low pH self-assembles after incubation at higher temperature into fibrils composed of antiparallel sheets that are stacked in a β2-type structure whose amide carbonyls have bifurcated H-bonds involving the side chains from the next sheet. Oligomers of Glu can also form such structures, and isotope labeling has provided insight into their out-of-register antiparallel structure [ Biomacromolecules 2013 , 14 , 3880 - 3891 ]. In this paper we report IR and VCD spectra and transmission electron micrograph (TEM) images for a series of alternately sequenced oligomers, Lys-(Aaa-Glu)5-Lys-NH2, where Aaa was varied over a variety of polar, aliphatic, or aromatic residues. Their spectral and TEM data show that these oligopeptides self-assemble into different structures, both local and morphological, that are dependent on both the nature of the Aaa side chains and growth conditions employed. Such alternate peptides substituted with small or polar residues, Ala and Thr, do not yield fibrils; but with β-branched aliphatic residues, Val and Ile, that could potentially pack with Glu side chains, these oligopeptides do show evidence of β2-stacking. By contrast, for Leu, with longer side chains, only β1-stacking is seen while with even larger Phe side chains, either β-form can be detected separately, depending on preparation conditions. These structures are dependent on high temperature incubation after reducing the pH and in some cases after sonication of initial fibril forms and reincubation. Some of these fibrillar peptides, but not all, show enhanced VCD, which can offer evidence for formation of long, multistrand, often twisted structures. Substitution of Glu with residues having selected side chains yields a variety of morphologies, leading to both β1- and β2-structures, that overall suggests two different packing modes for the hydrophobic side chains depending on size and type.

Isomorphous Crystals from Diynes and Bromodiynes Involved in Hydrogen and Halogen Bonds

Isomorphous crystals of two diacetylene derivatives with carbamate functionality (BocNH-CH2-diyne-X, where X = H or Br) have been obtained. The main feature of these structures is the original 2D arrangement (as supramolecular sheets or walls) in which the H bond and halogen bond have a prominent effect on the whole architecture. The two diacetylene compounds harbor neighboring carbamate (Boc protected amine) and conjugated alkyne functionalities. They differ only by the nature of the atom located at the penultimate position of the diyne moiety, either a hydrogen atom or a bromine atom. Both of them adopt very similar 2D wall organizations with antiparallel carbamates (as in antiparallel beta pleated sheets). Additional weak interactions inside the same walls between molecular bricks are H bond interactions (diyne-H···O=C) or halogen bond interactions (diyne-Br···O=C), respectively. Based on crystallographic atom coordinates, DFT (B3LYP/6-31++G(d,p)) and DFT (M06-2X/6-31++G(d,p)) calculations were performed on these isostructural crystals to gain insight into the intermolecular interactions.

The molecular organization of the beta-sheet region in Corneous beta-proteins (beta-keratins) of sauropsids explains its stability and polymerization into filaments.

The hard corneous material of avian and reptilian scales, claws, beak and feathers is mainly derived from the presence of proteins formerly known as beta-keratins but now termed Corneous beta-proteins of sauropsids to distinguish them from keratins, which are members of the intermediate filament protein family. The modeling of the conserved 34 amino acid residues long central beta-sheet region of Corneous beta-proteins using an ab initio protein folding and structure prediction algorithm indicates that this region is formed by four antiparallel beta-sheets. Molecular dynamic simulations and Molecular Mechanics/Poisson Boltzmann Surface Area (MM-PBSA) analysis showed that the disposition of polar and apolar amino acids within the beta-region gives rise to an amphipathic core whose stability is further increased, especially in an aqueous environment, by the association into a dimer due to apolar interactions and specific amino-acid interactions. The dimers in turn polymerize into a 3nm thick linear beta-filament due to van der Waals and hydrogen-bond interactions. It is suggested that once this nuclear core of anti-parallel sheets evolved in the genome of a reptilian ancestor of the extant reptiles and birds about 300 millions years ago, new properties emerged in the corneous material forming scales, claws, beaks and feathers in these amniotes based on the tendency of these unique corneous proteins to form stable filaments different from keratin intermediate filaments or sterical structures formed by other corneous proteins so far known.

Crystal growth and physical properties of the organic salt benzimidazolium 3-nitrophthalate

From the point of view of crystal growth and design, it is interesting to study the effect of different isomers on the resulting structure. Single crystals of the organic salt benzimidazolium 3-nitrophthalate (BZD+·mNPA−) have been grown by slow evaporation from methanol solution. Single crystal X-ray diffraction demonstrated that the crystal structure belongs to the monoclinic system with space group P21/c. The properties of the functional groups have been studied by FTIR spectroscopy, FT-Raman spectral analysis and 1H and 13C NMR spectroscopy. The emission spectra of BZD+·mNPA− were recorded in different solvents; they exhibit a negative solvatochromatic effect when the polarity of the solvent is increased. The thermal stability, dielectric constant and dielectric loss have been investigated and the HOMO and LUMO energy levels have been calculated using density functional theory (DFT). The crystal structure of BZD+·mNPA− has been compared with that of benzimidazolium 2-nitroterephthalate (BZD+·NTPh−). The interactions of the isomers 3-nitrophthalate or 2-nitroterephthalate salts with benzimidazolium are very similar; however, the different positions of the carboxyl groups on the benzene rings changes the layout of the structure, which leads to undulating sheets for 3-nitrophthalate and flat antiparallel sheets for 2-nitrotherephthalate. The salt decomposes at around 180 °C.

Diversification of β-Augmentation Interactions between CDI Toxin/Immunity Proteins

Contact-dependent growth inhibition (CDI) is a widespread mechanism of inter-bacterial competition mediated by the CdiB/CdiA family of two-partner secretion proteins. CdiA effectors carry diverse C-terminal toxin domains (CdiA-CT), which are delivered into neighboring target cells to inhibit growth. CDI+ bacteria also produce CdiI immunity proteins that bind specifically to cognate CdiA-CT toxins and protect the cell from auto-inhibition. Here, we compare the structures of homologous CdiA-CT/CdiI complexes from Escherichia coli EC869 and Yersinia pseudotuberculosis YPIII to explore the evolution of CDI toxin/immunity protein interactions. Both complexes share an unusual β-augmentation interaction, in which the toxin domain extends a β-hairpin into the immunity protein to complete a six-stranded anti-parallel sheet. However, the specific contacts differ substantially between the two complexes. The EC869 β-hairpin interacts mainly through direct H-bond and ion-pair interactions, whereas the YPIII β-hairpin pocket contains more hydrophobic contacts and a network of bridging water molecules. In accord with these differences, we find that each CdiI protein only protects target bacteria from its cognate CdiA-CT toxin. The compact β-hairpin binding pocket within the immunity protein represents a tractable system for the rationale design of small molecules to block CdiA-CT/CdiI complex formation. We synthesized a macrocyclic peptide mimic of the β-hairpin from EC869 toxin and solved its structure in complex with cognate immunity protein. These latter studies suggest that small molecules could potentially be used to disrupt CDI toxin/immunity complexes.

Crystal structure and physical properties of Antheraea yamamai silk fibers: Long poly(alanine) sequences are partially in the crystalline region

Antheraea yamamai silk contains long poly(alanine) sequences that contribute to β-sheet crystals, which are responsible for the high tensile strength of silk fibers. We determined the crystal structure, physical properties, and morphology of A. yamamai cocoon silk fibers. The crystal lattice of A. yamamai silk fiber consists of antiparallel β-pleated sheets with an orthogonal unit cell (a = 10.72 Å, b = 9.73 Å, c [fiber axis] = 6.80 ± 0.05 Å). Wide-angle X-ray scattering and birefringence measurements revealed that stretching deformation does not affect the crystal structure but contributes to alignment of silk molecules along the fiber axis in the partial amorphous phase. Crystallinity and amino acid sequence analyses suggested that the poly(alanine) region can be partially crystallized. The long poly(alanine) sequences therefore do not contribute to the mechanical, thermal, or structural properties of A. yamamai silk. Our results may aid the design and development of A. yamamai silk-based materials.