Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
9
pubmed:dateCreated
1993-7-7
pubmed:abstractText
Based on recent X-ray structural information, six site-directed mutants of human cyclophilin A (hCyPA) involving residues in the putative active site--H54, R55, F60, Q111, F113, and H126--have been constructed, overexpressed, and purified from Escherichia coli to homogeneity. The proteins W121A (Liu, J., Chen, C.-M., & Walsh, C.T., 1991a, Biochemistry 30, 2306-2310), H54Q, R55A, F60A, Q111A, F113A, and H126Q were assayed for cis-trans peptidyl-prolyl isomerase (PPIase) activity, their ability to bind the immunosuppressive drug cyclosporin A (CsA), and protein phosphatase 2B (calcineurin) inhibition in the presence of CsA. Results indicate that H54Q, Q111A, F113A, and W121A retain 3-15% of the catalytic efficiency (kcat/Km) of wild-type recombinant hCyPA. The remaining three mutants (R55A, F60A, and H126Q) each retain less than 1% of the wild-type catalytic efficiency, indicating participation by these residues in PPIase catalysis. Each of the mutants bound to a CsA affinity matrix. The mutants R55A, F60A, F113A, and H126Q inhibited calcineurin in the presence of CsA, whereas W121A did not. Although CsA is a competitive inhibitor of PPIase activity, it can complex with enzymatically inactive cyclophilins and inhibit the phosphatase activity of calcineurin.
pubmed:grant
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-1315036, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-1538777, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-1652374, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-1709302, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-1715244, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-1715516, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-1825312, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-1896075, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-1946361, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2001362, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2007139, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2054355, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2054356, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2059621, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2111742, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2179953, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2184885, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2477714, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2477715, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2492638, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2531848, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2595372, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-2644542, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-3013843, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-3881765, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-502865, http://linkedlifedata.com/resource/pubmed/commentcorrection/1338979-6238408
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0961-8368
pubmed:author
pubmed:issnType
Print
pubmed:volume
1
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1092-9
pubmed:dateRevised
2010-9-7
pubmed:meshHeading
pubmed-meshheading:1338979-Amino Acid Isomerases, pubmed-meshheading:1338979-Amino Acid Sequence, pubmed-meshheading:1338979-Base Sequence, pubmed-meshheading:1338979-Binding Sites, pubmed-meshheading:1338979-Calcineurin, pubmed-meshheading:1338979-Calmodulin-Binding Proteins, pubmed-meshheading:1338979-Carrier Proteins, pubmed-meshheading:1338979-Circular Dichroism, pubmed-meshheading:1338979-Cyclosporine, pubmed-meshheading:1338979-Humans, pubmed-meshheading:1338979-Kinetics, pubmed-meshheading:1338979-Molecular Sequence Data, pubmed-meshheading:1338979-Mutagenesis, Site-Directed, pubmed-meshheading:1338979-Oligodeoxyribonucleotides, pubmed-meshheading:1338979-Peptidylprolyl Isomerase, pubmed-meshheading:1338979-Phosphoprotein Phosphatases, pubmed-meshheading:1338979-Protein Binding, pubmed-meshheading:1338979-Protein Conformation, pubmed-meshheading:1338979-Protein Structure, Secondary, pubmed-meshheading:1338979-Recombinant Proteins, pubmed-meshheading:1338979-Spectrometry, Fluorescence
pubmed:year
1992
pubmed:articleTitle
Active site mutants of human cyclophilin A separate peptidyl-prolyl isomerase activity from cyclosporin A binding and calcineurin inhibition.
pubmed:affiliation
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.