Interaction of isoforms of S100 protein with smooth muscle caldesmon.
Interaction of S100a and S100b with duck gizzard caldesmon was investigated by means of native gel electrophoresis, fluorescent spectroscopy and disulfide crosslinking. Both isoforms of S100 interact with intact caldesmon and its C-terminal deletion mutant 606C (residues 606-756) with apparent Kd of 0.2-0.6 microM thus indicating that the S100-binding site is located in the C-terminal domain of caldesmon. The single SH group of duck gizzard caldesmon can be crosslinked to Cys-84 of the beta-chain or to Cys-85 of the alpha-chain of S100. Crosslinking of S100 reduces the inhibitory action of caldesmon on actomyosin ATPase activity. S100 reverses the inhibitory action of intact caldesmon and its deletion mutants 606C (residues 606-756) and H9 (residues 669-737) as effectively as calmodulin. S100a has higher affinity to caldesmon and is more effective than S100b in reversing caldesmon-induced inhibition of actomyosin ATPase activity. Although monomeric (calmodulin, troponin C) and dimeric (S100) Ca-binding proteins have different sizes and structures they interact with caldesmon in a very similar fashion.
Polyakov
A A
AA
Department of Biochemistry, School of Biology, Moscow State University, Russia.
Huber
P A
PA
Marston
S B
SB
Gusev
N B
NB
D000205:Actomyosin / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D015415:Biomarkers; D001921:Brain / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D019281:Dimerization; D004372:Ducks; D005895:Gizzard, Avian; D007700:Kinetics; D009130:Muscle, Smooth / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009414:Nerve Growth Factors; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins / Q000737:chemistry* / Q000378:metabolism*; D017384:Sequence Deletion
D015415:Biomarkers; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D009414:Nerve Growth Factors; D011994:Recombinant Proteins; C030564:S-100 calcium-binding protein alpha subunit; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; D000205:Actomyosin; D000251:Adenosine Triphosphatases