Site-specific conjugation of a temperature-sensitive polymer to a genetically-engineered protein.

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Chilkoti, A; Chen, G; Stayton, PS; Hoffman, AS

Published Date

  • November 1994

Published In

Volume / Issue

  • 5 / 6

Start / End Page

  • 504 - 507

PubMed ID

  • 7873655

Electronic International Standard Serial Number (EISSN)

  • 1520-4812

International Standard Serial Number (ISSN)

  • 1043-1802

Digital Object Identifier (DOI)

  • 10.1021/bc00030a004


  • eng