Detection of binding partners to the profilin:actin complex by far Western and mass spectrometry analyses.
A method to search for interaction partners to the profilin:actin complex that can distinguish molecules that preferentially bind the complex from those that interact with profilin or actin separately is described. The procedure should be applicable for any situation where cell extracts or other complex samples are screened for the presence of protein molecules specifically recognizing a protein complex but having no or low affinity for its individual components. The method is readily combined with mass spectrometry for direct identification of detected proteins. In this study, Mena and Hsp70 were detected as interaction partners to profilin:actin.
Johansson
Thomas
T
Department of Cell Biology, WGI, Stockholm University, S-106 91 Stockholm, Sweden.
Grenklo
Staffan
S
Karlsson
Roger
R
D000199:Actins / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002457:Cell Extracts / Q000032:analysis; D003285:Contractile Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000032:analysis / Q000378:metabolism*; D051304:Profilins; D011485:Protein Binding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013950:Thymus Gland / Q000737:chemistry
D000199:Actins; D002457:Cell Extracts; D003285:Contractile Proteins; D008840:Microfilament Proteins; D046912:Multiprotein Complexes; C495738:PFN1 protein, human; D051304:Profilins