Purification of a 47-kDa calmodulin-binding polypeptide as an actin-binding protein from Neurospora crassa.
We have enriched a 47-kDa polypeptide (p47) from Neurospora crassa on the basis of its affinity to calmodulin. The p47 was purified to homogeneity by chromatography on a Mono S cation exchange column and evidence is presented that the polypeptide co-sediments specifically with F-actin. The intracellular distribution of p47 and actin was also examined using indirect double immunofluorescence staining of cells at different stages of development. Our results suggest that by altering the conformation binding site of actin to p47, calmodulin could play a regulatory role in the polarized hyphal growth of N. crassa.
Capelli
N
N
Laboratory of Biochemistry and Plant Physiology, University of Geneva, Switzerland.
Barja
F
F
van Tuinen
D
D
Monnat
J
J
Turian
G
G
Ortega Perez
R
R
D000199:Actins / Q000276:immunology / Q000378:metabolism*; D001665:Binding Sites; D015153:Blotting, Western; D002147:Calmodulin / Q000378:metabolism*; D002847:Chromatography, Agarose; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D019084:Fluorescent Antibody Technique, Indirect; D009492:Neurospora crassa / Q000737:chemistry*; D010455:Peptides / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism*; D011485:Protein Binding
D000199:Actins; D002147:Calmodulin; D010455:Peptides