A role in enzyme activation for the N-terminal leader sequence in calmodulin.
We have found that deletion of residues 2-8 from the N-terminal leader sequence: Ala1-Asp2-Gln-Leu4-Thr-Glu6-Glu-Gln8, in calmodulin abolishes calmodulin-dependent activation of skeletal muscle myosin light chain kinase activity and reduces calmodulin-dependent activation of smooth muscle myosin light chain kinase activity to approximately 50% of the maximum level measured at a saturating calmodulin concentration. Calmodulin-dependent activation of cerebellar nitric oxide synthase activity is not affected by this deletion. Overlapping tripeptide deletions from the leader sequence indicate that the acidic cluster, Glu6-Glu-Gln8, contains the determinants necessary for activation of myosin light chain kinase activity. Deletion of Asp2-Gln-Leu4 has no effect on activation of enzyme activity. Based on enzyme kinetic analyses, deletions in the leader sequence have little or no effect on the apparent affinities of calmodulin for the synthase or the two kinases. Since the N-terminal leader does not appear to play a significant structural role in the complexes between calmodulin and peptides representing the calmodulin-binding domains in the two kinases, our results indicate that it participates in secondary interactions with these enzymes that are important to activation, but not to recognition or binding of calmodulin.
Persechini
A
A
Department of Pharmacology and Physiology, University of Rochester Medical Center, Rochester, New York 14642, USA.
Gansz
K J
KJ
Paresi
R J
RJ
D000595:Amino Acid Sequence; D000818:Animals; D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism*; D004789:Enzyme Activation; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000201:enzymology / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D021382:Protein Sorting Signals / Q000737:chemistry*; D011817:Rabbits; D017384:Sequence Deletion
D002147:Calmodulin; D021382:Protein Sorting Signals; D019001:Nitric Oxide Synthase; D009219:Myosin-Light-Chain Kinase