Dataset Information

MS characterization of spidroin-1

ABSTRACT: For MS characterization of major ampullate spidroin-1 from the three species of Nephila clavipes, edulis and madagascariensis, an experimental approach was developed combining 2-DE with multiple proteolytic in-gel digestion (trypsin, chymotrypsin, pepsin, proteinase K, subtilisin and proteinase 10) followed by NanoLC and mass spectrometry analysis using collision-induced dissociation (CID) and electron-transfer dissociation (ETD) for fragmentation. MASCOT searches were done by using the MASCOT 2.2.06 (Matrix Science, London, UK) against database for protein identification and post-translational modifications.

INSTRUMENT(S): Bruker Daltonics HCT Series, instrument model

ORGANISM(S): Nephila Clavipes (golden Silk Orbweaver)

SUBMITTER: José Roberto Aparecido dos Santos-Pinto

LAB HEAD: José Roberto Aparecido dos Santos Pinto

PROVIDER: PXD000057 | Pride | 2022-02-22

REPOSITORIES: Pride

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Publications

Structural Model for the Spider Silk Protein Spidroin-1.

dos Santos-Pinto José Roberto Aparecido JR Arcuri Helen Andrade HA Priewalder Helga H Salles Heliana Clara HC Palma Mario Sergio MS Lubec Gert G

Journal of proteome research 20150811 9

Most reports about the 3-D structure of spidroin-1 have been proposed for the protein in solid state or for individual domains of these proteins. A gel-based mass spectrometry strategy using collision-induced dissociation (CID) and electron-transfer dissociation (ETD) fragmentation methods was used to completely sequence spidroins-1A and -1B and to assign a series of post-translational modifications (PTMs) on to the spidroin sequences. A total of 15 and 16 phosphorylation sites were detected on ...[more]

PMID: 26211688

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Project description:Marine biomonitoring programs in the U.S. and Europe have historically relied on monitoring tissue concentrations of bivalves to monitor contaminant levels and ecosystem health. By integrating M-bM-^@M-^Xomic methods with these tissue residue approaches we can uncover mechanistic insight to help link tissue concentrations to toxic effects. In an effort to identify novel biomarkers and better understand the molecular toxicology of metal bioaccumulation in bivalves, we exposed the blue mussel, Mytilus edulis L., to sub-lethal concentrations (0.54 M-oM-^AM--M) of cadmium, lead, and a Cd+Pb mixture. Metal concentrations were measured in gill tissues at 1, 2, and 4 weeks, and increased linearly over the 4 week duration. In addition, there was evidence that Pb interfered with Cd uptake in the mixture treatment. Using a 3025 sequence microarray for M. edulis, we performed transcriptomic analysis, identifying 57 differentially expressed sequences. Hierarchical clustering of these sequences successfully distinguished the different treatment groups demonstrating that the expression profiles were reproducible among the treatments. Enrichment analysis of gene ontology terms identified several biological processes that were perturbed by the treatments, including nucleoside phosphate biosynthetic processes, mRNA metabolic processes, and response to stress. To identify transcripts whose expression level correlated with metal bioaccumulation, we performed Pearson correlation analysis. Several transcripts correlated with gill metal concentrations including mt10, mt20, and contig 48, an unknown transcript containing a wsc domain. In addition, three transcripts directly involved in the Unfolded protein response (UPR) were induced in the metal treatments at 2 weeks and were further up-regulated at 4 weeks. Overall, correlation of tissue concentrations and gene expression responses indicates that as mussels accumulate higher concentrations of metals, initial stress responses are mobilized to protect tissues. However, given the role of UPR in apoptosis, it serves as an early indicator of stress, which once overwhelmed will result in adverse physiological effects. A total of 52 samples including initial samples and 3 time-points following exposure to cadmium, lead, or a mixture. There were four replicate samples for each treatment and time-point. An additional three replicate samples were used for RT-qPCR.

Dataset Information

MS characterization of spidroin-1

Publications

Structural Model for the Spider Silk Protein Spidroin-1.

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