Functional and structural similarities between the inhibitory region of troponin I coded by exon VII and the calmodulin-binding regulatory region of the catalytic subunit of phosphorylase kinase.
A sequence homology has been noted between the carboxyl quarter of the catalytic gamma subunit of phosphorylase kinase and the region of troponin I coded by exon VII. Because this portion of troponin I contains the inhibitory region that interacts with actin and troponin C, we have examined whether the gamma subunit of phosphorylase kinase can functionally mimic troponin I by also interacting with actin and troponin C. We have found that troponin C not only activates the isolated gamma subunit of phosphorylase kinase but also binds with approximately the same affinity as calmodulin. Although actin had no effect on the activity of the gamma subunit alone, it did inhibit the activity of gamma-calmodulin and gamma-troponin C complexes. Conversely, the gamma subunit was able to inhibit actomyosin ATPase in a process that could be overcome by calmodulin. These results suggest that actin and calmodulin (or troponin C) compete for binding to the gamma subunit. Moreover, the structural and functional similarities between the gamma subunit and troponin I suggest that the gamma subunit of phosphorylase kinase may have evolved from the fusion of a protein kinase protogene with a progenitor of exon VII of troponin I.
Paudel
H K
HK
Department of Biochemistry, College of Medicine, University of Tennessee, Memphis 38163.
Carlson
G M
GM
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002147:Calmodulin / Q000378:metabolism*; D002417:Cattle; D005091:Exons; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology / Q000378:metabolism; D009218:Myosins / Q000378:metabolism; D010764:Phosphorylase Kinase / Q000235:genetics* / Q000378:metabolism; D011817:Rabbits; D012689:Sequence Homology, Nucleic Acid; D014336:Troponin / Q000235:genetics* / Q000378:metabolism; D019210:Troponin I
D000199:Actins; D002147:Calmodulin; D046911:Macromolecular Substances; D014336:Troponin; D019210:Troponin I; D010764:Phosphorylase Kinase; D009218:Myosins