A METHOD FOR THE SAMPLE HANDLING AND ANALYSIS OF BIO-ACTIVE PEPTIDES

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Authors
  1. Hancock, J.R.
  2. Provost, L.R.
  3. D'Agostino, P.A.
  4. Toernes, J.A.
Corporate Authors
Defence Research Establishment Suffield, Ralston ALTA (CAN);Norwegian Defence Research Establishment, Kjeller (Norway)
Abstract
A method has been developed for the sample handling and analysis of bio-active peptides. Based on electrospray ionization mass spectrometry (ESMI-MS), the method is applicable to the target analysis of known peptides. ESI-MS is used to determine the molecular mass of the peptide and provisional identification is based on matching the molecular mass with that of known peptides in a database. Disulfide bridge reductive alkylation is used to determine the number of cysteines in the peptide as well as the presence of disulfide bridges. The peptide is then enzymatically digested and the digestion products analyzed by ESI-MS. The resulting massmap is compared to the masses predicted from the structure of the peptide. Finally, the peptide and its enzymatic fragments are analyzed by liquid chromatography (LC) ESI-MS using collisional activated disassociation (CAD) conditions which promote the formation of product ions from which it is possible to determine the amino acid sequence of the peptide. The developed method was applied to the analysis and identification of alpha-conotoxin GI. The monisotopic molecular mass was determined to be 1436.42 + or - 0.13 u (n=6). An search of the in-house database determined that the only match within one mass unit of the calculated molecular mass was alpha-conotoxin GI with a theoretical molecular mass of 1436.48 u. TRUNCATED
Keywords
Bioactive peptides;Toxic peptides;Electrospray;Collisional Activated Disassociation;Disulfide Bridge Reduction;Conotoxin
Report Number
DRES-699 —
Date of publication
01 May 1998
Number of Pages
23
DSTKIM No
98-02056
CANDIS No
508814
Format(s):
Hardcopy;Document Image stored on Optical Disk

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