Immunological and biochemical studies on the relationship between two actin-binding proteins, phosphofructokinase and gelsolin.
Phosphofructokinase and gelsolin-like proteins coexist in many muscle and non-muscle tissues. They are both actin-binding proteins, and some of their biochemical parameters are remarkably similar. In a previous report [Füchtbauer, A., Jockusch, B. M., Leberer, E. & Pette, D. (1986) Proc. Natl Acad. Sci. USA 83, 9502-9506] it was shown that phosphofructokinase preparations contained actin-filament-severin activities characteristic for gelsolin. Therefore, we investigated a possible relationship between these proteins with respect to their actin-binding properties. Immunoblotting experiments with specific and non-cross-reacting antibodies to both proteins revealed two distinct polypeptides with slightly different molecular mass in SDS-PAGE of crude extracts from rabbit skeletal muscle, indicating that phosphofructokinase and gelsolin are not identical. An actin-filament-severing activity as well as the component detected by anti-gelsolin were found to copurify with phosphofructokinase during its preparation. However, the presumptive gelsolin was completely eliminated after a heat-denaturation step leaving the phosphofructokinase activity unaffected. Purified phosphofructokinase had no effects on the polymer state of preformed actin filaments. Unlike gelsolin, phosphofructokinase did not promote nucleation of actin polymerization but delayed the nucleation step. We therefore conclude that phosphofructokinase and gelsolin are functionally and structurally distinct proteins.
Buss
F
F
Developmental Biology Unit, University of Bielefeld, Federal Republic of Germany.
Hinssen
H
H
Jockusch
B M
BM
D000199:Actins / Q000378:metabolism*; D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex / Q000032:analysis; D002135:Calcium-Binding Proteins / Q000276:immunology / Q000378:metabolism*; D003429:Cross Reactions; D005753:Gastric Mucosa / Q000378:metabolism; D018260:Gelsolin; D007700:Kinetics; D008840:Microfilament Proteins / Q000276:immunology / Q000378:metabolism*; D009130:Muscle, Smooth / Q000378:metabolism; D009132:Muscles / Q000201:enzymology / Q000378:metabolism; D010732:Phosphofructokinase-1 / Q000276:immunology / Q000378:metabolism*; D011485:Protein Binding; D011817:Rabbits; D013552:Swine
D000199:Actins; D000906:Antibodies; D000936:Antigen-Antibody Complex; D002135:Calcium-Binding Proteins; D018260:Gelsolin; D008840:Microfilament Proteins; D010732:Phosphofructokinase-1