SITE-SPECIFIC CONJUGATION OF A TEMPERATURE-SENSITIVE POLYMER TO A GENETICALLY-ENGINEERED PROTEIN

被引：117

作者：

CHILKOTI, A ^{[1
]}

CHEN, GH ^{[1
]}

STAYTON, PS ^{[1
]}

HOFFMAN, AS ^{[1
]}

机构：

[1] UNIV WASHINGTON,CTR BIOENGN,SEATTLE,WA 98195

来源：

BIOCONJUGATE CHEMISTRY | 1994年 / 5卷 / 06期

关键词：

D O I：

10.1021/bc00030a004

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A genetically-engineered mutant of cytochrome b5, incorporating a unique cysteine residue, was conjugated to maleimide-terminated oligo(N-isopropylacrylamide). The conjugation of the protein by reaction of the cysteine residue, precisely positioned by site-directed mutagenesis techniques, with an activated oligomer containing only one reactive end group in the oligomer chain permits the site-specific and stoichiometric conjugation of the oligomer with the protein. The protein-oligomer conjugate was shown to exhibit lower critical solution temperature (LCST) behavior, similar to the free oligomer. Furthermore, the LCST behavior of the protein-oligomer conjugate is reversible and allows selective precipitation of the conjugate above its LCST.

引用

页码：504 / 507

页数：4

共 17 条

[1]

CHEN GH, 1994, J BIOMAT SCI-POLYM E, V5, P371

[2] PREPARATION AND PROPERTIES OF THERMOREVERSIBLE, PHASE-SEPARATING ENZYME-OLIGO(N-ISOPROPYLACRYLAMIDE) CONJUGATES [J].