Document Detail


Enhancing the stability of ¹⁸O-labeled peptides through removal of immobilized trypsin by ZipTips.
MedLine Citation:
PMID:  20816659     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
Trypsin-catalyzed ¹⁸O labeling is increasingly used in shotgun proteomics for relative peptide/protein quantitation. However, precise quantitative measurements are often complicated by the instability of ¹⁸O-labeled peptides caused mainly by oxygen back-exchange. Although a number of attempts have been made to reduce or prevent oxygen back-exchange, there is still room for improvement. Here we demonstrate that the removal of immobilized trypsin by filtration using ZipTips can efficiently minimize oxygen back-exchange and enhance the stability of ¹⁸O-labeled peptides under various pH conditions. The ¹⁸O-labeled peptides processed by the approach were successfully separated by immobilized pH gradient-isoelectric focusing (IPG-IEF), and no marked decrease in the extent of labeling was observed. The results also demonstrated that there was no correlation between the extent of ¹⁸O labeling and molecular weight or isoelectric point (pI). The approach presented here is especially applicable to microscale samples. Its ability to generate stably ¹⁸O-labeled samples without back-exchange should expand the application scope of the ¹⁸O-labeling technique.
Authors:
Mao-Yu Li; Fang Peng; Jian-Hong Zuo; Hong Yi; Can-E Tang; Cui Li; Peng-Fei Zhang; Zhu-Chu Chen; Zhi-Qiang Xiao
Related Documents :
8678299 - Purification of synthetic peptide libraries by affinity chromatography using the avidin...
16038319 - Use of mixed-mode sorbents for the electrochromatographic separation of thrombin recept...
7174669 - Human hypoxanthine-guanine phosphoribosyltransferase. tryptic peptides and post-transla...
11100869 - Influence of the structure of cyclodextrins and amino acid sequence of dipeptides and t...
16348979 - Long-term production of ergot peptides by immobilized claviceps purpurea in semicontinu...
19928919 - Peptide fractionation and free radical scavenging activity of zein hydrolysate.
24278589 - The safety evaluation of a potent angiogenic activator, synthetic peptide (sfklry-nh2) ...
23895279 - A novel form of bacterial resistance to the action of eukaryotic host defence peptides,...
2023899 - Isolation and characterization of two peptides with prolactin release-inhibiting activi...
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-09-20
Journal Detail:
Title:  Analytical biochemistry     Volume:  408     ISSN:  1096-0309     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2010-10-26     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  37-45     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier Inc. All rights reserved.
Affiliation:
Xiangya Hospital, Central South University, Changsha, China.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Usefulness of various MRI sequences in the diagnosis of viral encephalitis.
Next Document:  Monitoring nonenzymatic glycation of human immunoglobulin G by methylglyoxal and glyoxal: A spectros...