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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): LTQ Orbitrap, Bruker Daltonics HCT Series, HCTultra ETD II

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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			Action	DRS
	F047365.yep	Other
	F047367.yep	Other
	F047368.yep	Other
	F047369.yep	Other
	F047371.yep	Other

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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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