In:
Rapid Communications in Mass Spectrometry, Wiley, Vol. 22, No. 20 ( 2008-10-30), p. 3313-3319
Abstract:
We have developed a new method to determine the N‐terminal amino acid sequences of proteins, regardless of whether their N‐termini are modified. This method consists of the following five steps: (1) reduction, S‐alkylation and guanidination for targeted proteins; (2) coupling of sulfo‐NHS‐SS‐biotin to N α ‐amino groups of proteins; (3) digestion of the modified proteins by an appropriate protease followed by oxidation with performic acid; (4) specific isolation of N‐terminal peptides from digests using DITC resins; (5) de novo sequence analysis of the N‐terminal peptides by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) using the CAF (chemically assisted fragmentation) method or tandem mass spectrometric (MS/MS) analysis according to unblocked or blocked peptides, respectively. By employing DITC resins instead of avidin resins used in our previous method (Yamaguchi et al ., Rapid Commun. Mass Spectrom. 2007; 21: 3329), it has been possible to isolate selectively N‐terminal peptides from proteins regardless of modification of N‐terminal amino acids. Here we propose a universal method for N‐terminal sequence analysis of proteins. Copyright © 2008 John Wiley & Sons, Ltd.
Type of Medium:
Online Resource
ISSN:
0951-4198
,
1097-0231
Language:
English
Publisher:
Wiley
Publication Date:
2008
detail.hit.zdb_id:
2002158-6
detail.hit.zdb_id:
58731-X
SSG:
11
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