Document Detail


Three-step purification of a fragment of the large immunophilin FKBP52.
MedLine Citation:
PMID:  10681043     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PPIases catalyze the interconversion of cis and trans isomers of peptidyl-prolyl (Xaa-Pro) bonds in peptide and protein substrates. The PPIase family comprises three subfamilies, two of which interact with immunosuppressant drugs and are therefore termed immunophilins. One subgroup of the immunophilins are the FK506 binding proteins (FKBPs). FKBPs of a relative molecular mass higher than 40 000 also display chaperone activity and are part of the multichaperone complex that Hsp90 forms with substrate proteins. Their function in this chaperone complex is still enigmatic. To further characterize the function of FKBP52 we want to analyze constructs of FKBP52-fragments. Here we describe a fast and effective three-step purification procedure for a fragment of FKBP52 with a relative molecular mass of 48000, termed FKBP52-123, consisting of affinity chromatography, anion-exchange column and gel-permeation chromatography. A yield of 1 mg pure protein per gram of cells was achieved.
Authors:
F Pirkl
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of chromatography. B, Biomedical sciences and applications     Volume:  737     ISSN:  1387-2273     ISO Abbreviation:  J. Chromatogr. B Biomed. Sci. Appl.     Publication Date:  2000 Jan 
Date Detail:
Created Date:  2000-03-01     Completed Date:  2000-03-01     Revised Date:  2000-12-18    
Medline Journal Info:
Nlm Unique ID:  9714109     Medline TA:  J Chromatogr B Biomed Sci Appl     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  71-6     Citation Subset:  IM    
Affiliation:
Institut für Organische Chemie und Biochemie. Technische Universität München, Garching, Germany. franziska.pirkl@ch.tum.de
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MeSH Terms
Descriptor/Qualifier:
Chromatography, Affinity
Chromatography, Gel
Chromatography, Ion Exchange
Electrophoresis, Polyacrylamide Gel
Immunophilins / chemistry*
Peptide Fragments / isolation & purification*
Recombinant Proteins / isolation & purification
Tacrolimus Binding Proteins
Chemical
Reg. No./Substance:
0/Peptide Fragments; 0/Recombinant Proteins; EC 5.2.1.-/Tacrolimus Binding Proteins; EC 5.2.1.8/Immunophilins

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