KINETIC-ANALYSIS OF BIOTINYLATION OF SPECIFIC RESIDUES OF PEPTIDES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

被引：10

作者：

KUROSKY, A

MILLER, BT

KNOCK, SL

机构：

[1] UNIV TEXAS,MED BRANCH,INST MARINE BIOMED,GALVESTON,TX 77555

[2] UNIV TEXAS,MED BRANCH,DEPT ANAT & NEUROSCI,GALVESTON,TX 77555

来源：

JOURNAL OF CHROMATOGRAPHY | 1993年 / 631卷 / 1-2期

关键词：

D O I：

10.1016/0021-9673(93)80534-F

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A procedure employing C18 reversed-phase high-performance liquid chromatography (HPLC) is described for evaluating the kinetics of biotinylation of specific residues of peptides after reaction with N-hydroxysuccinimide esters of biotin. Utilizing this HPLC method, we determined the observed pseudo-first-order reaction mte constant (k1') of biotinylation of lysyl residues in two model peptides, [biotinyl-Ser108]ProA-egg laying hormone (108-121) and pGlu-Lys-Trp-Ala-Pro, to be 1.22 . 10(-2) s-1 and 1.08 . 10(-2) s-1, respectively, in 0.05 M sodium phosphate buffer, pH 8.2, at 25-degrees-C. The respective reaction half-lives of the two peptides were 57 s and 64 s. In addition, HPLC analytical methods were established for determining the time-course of hydrolysis of biotinylating reagents at acidic and alkaline pH and for evaluating biotin reagent homogeneity.

引用

页码：281 / 287

页数：7

共 9 条

[1]

Fields R, 1972, Methods Enzymol, V25, P464, DOI 10.1016/S0076-6879(72)25042-X

[2]

KNOCK SL, 1991, J BIOL CHEM, V266, P24413

[3] SOLID PHASE PEPTIDE SYNTHESIS .1. SYNTHESIS OF A TETRAPEPTIDE [J].