Photoaffinity Heterobifunctional Cross-Linking Reagents Based on N-(Azidobenzoyl)tyrosines

被引：9

作者：

Imai, Nobuyuki ^{[1
]}

Kometani, Tadashi ^{[1
]}

Crocker, Peter J. ^{[1
]}

Bowdan, Jean B. ^{[1
]}

Demir, Ayhan ^{[1
]}

Dwyer, Lori D. ^{[2
]}

Mann, Dennis M. ^{[2
]}

Vanaman, Thomas C. ^{[2
]}

Watt, David S. ^{[1
]}

机构：

[1] Univ Kentucky, Dept Chem, Lexington, KY 40506 USA

[2] Univ Kentucky, Dept Biochem, Lexington, KY 40506 USA

来源：

BIOCONJUGATE CHEMISTRY | 1990年 / 1卷 / 02期

基金：

美国国家科学基金会;

关键词：

22;

D O I：

10.1021/bc00002a008

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

New heterobifunctional cross-linking reagents that possessed a photoactive terminus, an electrophilic terminus, and a linking arm between the two termini that had a radiolabeled, enzymatically cleavable bond were synthesized. In a model study, succinimidyl N-[N'-(4-azidobenzoyl)tyrosyl]-beta-alanate (16A) was coupled to n-butylamine (a Lys surrogate), iodinated, and cleaved with chymotrypsin in the presence of tyrosylamide to afford the desired adduct N-(N'-(4-azidobenzoyl)-3-iodotyrosyl)tyrosinamide, thereby demonstrating the feasibility of the enzymatic cleavage. In a biochemical study, succinimidyl N-[N'-(3-azido-5-nitrobenzoyl)tyrosyl]-beta-alanate (16C) was coupled to Lys-75 of calmodulin (CaM), and the radioiodinated monoadduct was successfully photo-cross-linked, in a calcium-dependent manner, to the human erythrocyte plasma membrane Ca(2+), Mg(2+)-ATPase and to a synthetic fragment (M13) containing the CaM-binding region of myosin light-chain kinase. In the latter case, densitometry readings indicated 20% cross-linking efficiency.

引用

页码：138 / 143

页数：6

共 22 条

[1] USE OF ESTERS OF N-HYDROXYSUCCINIMIDE IN PEPTIDE SYNTHESIS [J].