Linked Life Data

8408423

Source:http://linkedlifedata.com/resource/pubmed/id/8408423

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1993-11-16
pubmed:abstractText
Several attempts to separate hydrophobic tryptic and cyanogen bromide-digested short peptides from Na,K-ATPase, using HPLC and different acid-organic solvent gradients, failed because of the insolubility of the peptides in the initial or final solvents of the gradients used for elution. Therefore, we opted to use a detergent-containing mobile phase. For sodium dodecyl sulphate-solubilized tryptic peptides of M(r) 7 x 10(3)-100 x 10(3), elution on a TSK-G3000SW size-exclusion column successfully separates families of peptides with a resolution of M(r) 5 x 10(3)-10 x 10(3). Peptides in these size ranges can then be resolved completely by tricine-sodium dodecyl sulphate gel electrophoresis, and identified by microsequencing after transfer to polyvinylidene difluoride paper. For separation of smaller peptides a Biosep-SEC-S2000 column, eluted at slow flow-rates, was evaluated. Use of ammonium chloride buffer allows sensitive detection at 214 nm. The separated fractions are resolved and identified on 16.5% tricine gels. Reasonable resolution has been obtained with defined cyanogen bromide fragments of myoglobin. Resolution of small tryptic and cyanogen bromide fragments of Na,K-ATPase is less successful, but the experiments suggest ways of improving the resolution of peptides in the range M(r) 2 x 10(3)-10 x 10(3).
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0021-9673
pubmed:author
pubmed:issnType
Print
pubmed:day
27
pubmed:volume
646
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
153-7
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed:year
1993
pubmed:articleTitle
Separation of membrane-embedded tryptic peptides of Na,K-ATPase by size-exclusion chromatography.
pubmed:affiliation
Department of Biochemistry, Weizmann Institute of Science, Rehovot, Israel.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.