Linked Life Data

9047315

Source:http://linkedlifedata.com/resource/pubmed/id/9047315

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
8
pubmed:dateCreated
1997-3-25
pubmed:abstractText
The regeneration of native recombinant hirudin variant 1 (rHV1) from the reduced unfolded form to the fully oxidized native state has been carried out with mixtures of oxidized and reduced dithiothreitol at pH 8.3 and 12 degrees C. The regeneration reaction was quenched at various times by the addition of 2-aminoethyl methanethiosulfonate to block unreacted sulfhydryl groups. The quenched protein-folding intermediates were fractionated by both capillary electrophoresis and a combination of anion exchange and reverse phase HPLC and characterized by mass spectrometry, amino acid analysis, and disulfide analysis. These intermediates (before quenching) were found to interconvert rapidly so as to achieve a steady-state distribution early in the regeneration process. The experimental data were fitted to a steady-state kinetic scheme. The analysis reveals that the rate-determining step in the regeneration of rHV1 with oxidized and reduced dithiothreitol involves the oxidation of one or more two-disulfide-containing species, most likely those already containing two native disulfide bonds. This regeneration mechanism is different from one that has been proposed by Chatrenet and Chang [(1993) J. Biol. Chem. 268, 20988]. The differences are discussed, and possible explanations for the differences are presented.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
36
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
2154-65
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1997
pubmed:articleTitle
Kinetic studies of the regeneration of recombinant hirudin variant 1 with oxidized and reduced dithiothreitol.
pubmed:affiliation
Baker Laboratory of Chemistry, Cornell University, Ithaca, New York 14853-1301, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't