A quantitative, high-throughput screen for protein stability.

Published

Journal Article

In proteomic research, it is often necessary to screen a large number of polypeptides for the presence of stable structure. Described here is a technique (referred to as SUPREX, stability of unpurified proteins from rates of H/D exchange) for measuring the stability of proteins in a rapid, high-throughput fashion. The method uses hydrogen exchange to estimate the stability of microgram quantities of unpurified protein extracts by using matrix-assisted laser desorption/ionization MS. The stabilities of maltose binding protein and monomeric lambda repressor variants determined by SUPREX agree well with stability data obtained from conventional CD denaturation of purified protein. The method also can detect the change in stability caused by the binding of maltose to maltose binding protein. The results demonstrate the precision of the method over a wide range of stabilities.

Full Text

Duke Authors

Cited Authors

  • Ghaemmaghami, S; Fitzgerald, MC; Oas, TG

Published Date

  • July 18, 2000

Published In

Volume / Issue

  • 97 / 15

Start / End Page

  • 8296 - 8301

PubMed ID

  • 10890887

Pubmed Central ID

  • 10890887

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.140111397

Language

  • eng

Conference Location

  • United States