BIOTINYLATED PEPTIDES PROTEINS .1. DETERMINATION OF STOICHIOMETRY OF DERIVATIZATION

被引：29

作者：

SMITH, JS

MILLER, BT

KNOCK, SL

KUROSKY, A

机构：

[1] UNIV TEXAS,MED BRANCH,DEPT HUMAN BIOL CHEM & GENET,GALVESTON,TX 77550

[2] UNIV TEXAS,MED BRANCH,DEPT ANAT & NEUROSCI,GALVESTON,TX 77550

[3] UNIV TEXAS,MED BRANCH,INST MARINE BIOMED,GALVESTON,TX 77550

来源：

ANALYTICAL BIOCHEMISTRY | 1991年 / 197卷 / 01期

关键词：

D O I：

10.1016/0003-2697(91)90384-6

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method is described for the determination of the stoichiometry of biotinylation of peptides and proteins after reaction with an N-hydroxysuccinimide ester of biotin containing the extended spacer arm 6-aminohexanoic acid (NHS-ε{lunate}Ahx-biotin). The method of analysis, based on the quantification of phenylthiocarbamyl derivatives of 6-aminohexanoic acid, is able to measure low picomolar amounts of biotinyl derivative. Analyses were performed using an automated on-line hydrolyzer-derivatizer followed by high-performance liquid chromatography. Compositional analyses determined for known peptides were in excellent agreement with analyses obtained by mass spectrometry. Procedures are also described for the production of biotinylated protein probes that can be labeled reproducibly to contain specific amounts of biotin. The analytical advantage and steric freedom provided by the 6-amino-hexanoic acid spacer arm argue strongly for the NHS-ε{lunate}Ahx-biotin reagent to be a reagent of choice for the biotinylation of peptides and proteins. © 1991.

引用

页码：247 / 253

页数：7

共 18 条

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