| Title: | Protein A-calmodulin fusions: a novel approach for investigating calmodulin function in yeast |
| Author(s): | Stirling DA; Petrie A; Pulford DJ; Paterson DT; Stark MJ; |
| Address: | "Department of Biochemistry, University of Dundee, UK" |
| DOI: | 10.1111/j.1365-2958.1992.tb01519.x |
| ISSN/ISBN: | 0950-382X (Print) 0950-382X (Linking) |
| Abstract: | "A novel gene fusion approach which may be of more general use has been developed for investigating the function of calmodulin in the budding yeast Saccharomyces cerevisiae. By fusing a portion of the Staphylococcus aureus spa gene (encoding protein A) to CMD1, the S. cerevisiae gene encoding calmodulin, we have generated a yeast calmodulin with an affinity tag able to bind immunoglobulins. The chimaeric protein A-calmodulin (ProtA-CaM) polypeptide functions in vivo and shows Ca(2+)-dependent binding to calmodulin target proteins. The spa-CMD1 fusion has been used (i) to prepare (by affinity chromatography) a fraction of yeast proteins which interact with calmodulin, (ii) to isolate genes encoding calmodulin target proteins by direct screening of an expression library, and (iii) to visualize calmodulin-binding proteins in crude extracts by Western blot analysis" |
| Keywords: | "Amino Acid Sequence Base Sequence Blotting, Western Calmodulin/*genetics/isolation & purification/metabolism Calmodulin-Binding Proteins/metabolism Cloning, Molecular DNA, Recombinant Gene Library Mating Factor Molecular Sequence Data Peptides/pharmacolog;" |
| Notes: | "MedlineStirling, D A Petrie, A Pulford, D J Paterson, D T Stark, M J eng Research Support, Non-U.S. Gov't England 1992/03/01 Mol Microbiol. 1992 Mar; 6(6):703-13. doi: 10.1111/j.1365-2958.1992.tb01519.x" |
