Abstract
The European freshwater mollusk Dreissena bugensis (quagga mussel), an invasive species to North America, adheres to surfaces underwater via the byssus: a non-living protein ‘anchor’. In spite of its importance as a biofouling species, the sequence of the majority of byssal proteins responsible for adhesion are not known, and little genomic data is available. To determine protein sequence information, we utilized next-generation RNA sequencing and de novo assembly to construct a cDNA library of the quagga mussel foot transcriptome, which contains over 200,000 transcripts. Quagga mussel byssal proteins were extracted from freshly induced secretions and analyzed using LC-MS/MS; peptide spectra were matched to the transcriptome to fingerprint the entire protein primary sequences. We present the full sequences of fourteen novel quagga mussel byssal proteins, named Dreissena bugensis foot proteins 4 to 17 (Dbfp4–Dbfp17), and new sequence data for two previously observed byssal proteins Dbfp1 and Dbfp2. Theoretical masses of the newly discovered proteins range from 4.3 kDa to 21.6 kDa. These protein sequences are unique but contain features similar to glue proteins from other species, including a high degree of polymorphism, proteins with repeated peptide motifs, disordered protein structure, and block structures.
Highlights
In contrast with freshwater mussels, the marine mussel adhesion system has been extensively studied[8,9,10]
In order to determine the primary sequence of quagga mussel byssal proteins, a bottom-up proteomics approach utilizing next-generation sequencing paired with LC-MS/MS protein identification was used. mRNA sequence data from three quagga mussel feet were pooled for de novo assembly using Trinity
The assembled library and sequence reads were deposited into the DNA Data Bank of Japan (DDBJ)
Summary
In contrast with freshwater mussels, the marine mussel adhesion system has been extensively studied[8,9,10]. In order to determine the primary sequence of byssal proteins, we paired massively parallel RNA sequencing technologies with high-resolution liquid chromatography tandem mass spectrometry (LC-MS/MS)[22]. This type of transcriptomic and proteomic analysis has emerged as a powerful method of analyzing the protein component of biological glues[22,23,24,25,26,27,28,29,30,31,32,33,34,35,36]. Using LC-MS/MS, extracted byssal proteins fragments were analyzed, and the resulting spectra were matched to the foot transcriptome library to fingerprint the protein primary structure for analysis
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