| Identification of cyclized calmodulin antagonists from a phage display random peptide library. | |
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MedLine Citation:
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PMID: 9237217 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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To isolate peptide ligands that bound calmodulin (CaM) specifically, we screened an M13 phage library displaying cyclized octamer random peptides with immobilized bovine CaM. Isolates were recovered, sequenced, and deduced to express nine independent peptides, five of which contained the sequence Trp-Gly-Lys (WGK). Four of the nine peptide sequences were synthesized in cyclized, biotinylated form. All of the peptides required Ca2+ to bind CaM. The cyclized, disulfide-bonded form of one such peptide, SCLRWGKWSNCGS, bound CaM better than its reduced form or an analogue in which the cysteine residues were replaced by serine. The cyclized peptide also exhibited the ability to inhibit CaM-dependent kinase activity. Systematic alanine substitution of residues in this peptide sequence implicate the tryptophan residue as being critical for binding, with other residues contributing to binding to varying degrees. Cloning of ligand targets (COLT) confirmed the specificity of one of the cyclized peptides, yielding full-length and C-terminal CaM clones, in addition to a full-length clone of troponin C, a CaM-related protein. This study has demonstrated that conformationally constrained peptides isolated from a phage library acted as specific, Ca(2+)-dependent CaM ligands. |
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Authors:
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H H Pierce; N Adey; B K Kay |
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Publication Detail:
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Type: In Vitro; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Molecular diversity Volume: 1 ISSN: 1381-1991 ISO Abbreviation: Mol. Divers. Publication Date: 1996 Aug |
Date Detail:
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Created Date: 1997-09-23 Completed Date: 1997-09-23 Revised Date: 2007-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9516534 Medline TA: Mol Divers Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 259-65 Citation Subset: IM |
Affiliation:
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Department of Biology, University of North Carolina at Chapel Hill 27599, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Animals Bacteriophage M13 / genetics Base Sequence Calcium-Calmodulin-Dependent Protein Kinases / antagonists & inhibitors Calmodulin / antagonists & inhibitors* Cattle Cloning, Molecular DNA, Complementary / genetics Directed Molecular Evolution / methods* Enzyme Inhibitors / chemistry, pharmacology Ligands Mice Peptide Library* Peptides, Cyclic / chemistry, genetics, pharmacology |
| Grant Support | |
ID/Acronym/Agency:
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T32 GM07092/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Calmodulin; 0/DNA, Complementary; 0/Enzyme Inhibitors; 0/Ligands; 0/Peptide Library; 0/Peptides, Cyclic; EC 2.7.11.17/Calcium-Calmodulin-Dependent Protein Kinases |
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