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Partial de novo sequencing and unusual CID fragmentation of a 7 kDa, disulfide-bridged toxin.

ABSTRACT: A 7 kDa toxin isolated from the venom of the Texas coral snake (Micrurus tener tener) was subjected to collision-induced dissociation (CID) and electron-transfer dissociation (ETD) analyses both before and after reduction at low pH. Manual and automated approaches to de novo sequencing are compared in detail. Manual de novo sequencing utilizing the combination of high accuracy CID and ETD data and an acid-related cleavage yielded the N-terminal half of the sequence from the reduced species. The intact polypeptide, containing 3 disulfide bridges produced a series of unusual fragments in ion trap CID experiments: abundant internal amino acid losses were detected, and also one of the disulfide-linkage positions could be determined from fragments formed by the cleavage of two bonds. In addition, internal and c-type fragments were also observed.

SUBMITTER: Medzihradszky KF

PROVIDER: S-EPMC4367482 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature

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Partial de novo sequencing and unusual CID fragmentation of a 7 kDa, disulfide-bridged toxin.

Medzihradszky Katalin F KF Bohlen Christopher J CJ

Journal of the American Society for Mass Spectrometry 20120216 5

A 7 kDa toxin isolated from the venom of the Texas coral snake (Micrurus tener tener) was subjected to collision-induced dissociation (CID) and electron-transfer dissociation (ETD) analyses both before and after reduction at low pH. Manual and automated approaches to de novo sequencing are compared in detail. Manual de novo sequencing utilizing the combination of high accuracy CID and ETD data and an acid-related cleavage yielded the N-terminal half of the sequence from the reduced species. The ...[more]

PMID: 22351294

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