Hypothesis for a serine proteinase-like domain at the COOH terminus of Slowpoke calcium-activated potassium channels.
Bovine pancreatic trypsin inhibitor (BPTI) is a 58-residue protein with three disulfide bonds that belongs to the Kunitz family of serine proteinase inhibitors. BPTI is an extremely potent inhibitor of trypsin, but it also specifically binds to various active and inactive serine proteinase homologs with KD values that range over eight orders of magnitude. We previously described an interaction of BPTI at an intracellular site that results in the production of discrete subconductance events in large conductance Ca2+ activated K+ channels (Moss, G.W.J., and E. Moczydlowski. 1996, J. Gen. Physiol, 107:47-68). In this paper, we summarize a variety of accumulated evidence which suggests that BPTI binds to a site on the KCa channel protein that structurally resembles a serine proteinase. One line of evidence includes the finding that the complex of BPTI and trypsin, in which the inhibitory loop of BPTI is masked by interaction with trypsin, is completely ineffective in the production of substate events in the KCa channel. To further investigate this notion, we performed a sequence analysis of the alpha-subunit of cloned slowpoke KCa channels from Drosophila and mammals. This analysis suggests that a region of approximately 250 residues near the COOH terminus of the KCa channel is homologous to members of the serine proteinase family, but is catalytically inactive because of various substitutions of key catalytic residues. The sequence analysis also predicts the location of a Ca(2+)-binding loop that is found in many serine proteinase enzymes. We hypothesize that this COOH-terminal domain of the slowpoke KCa channel adopts the characteristic double-barrel fold of serine proteinases, is involved in Ca(2+)-activation of the channel, and may also bind other intracellular components that regulate KCa channel activity.
Moss
G W
GW
Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520-8066, USA.
Marshall
J
J
Moczydlowski
E
E
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000502:physiology*; D006801:Humans; D008957:Models, Genetic; D008969:Molecular Sequence Data; D015221:Potassium Channels / Q000235:genetics*; D012697:Serine Endopeptidases / Q000235:genetics*
D015221:Potassium Channels; D012697:Serine Endopeptidases; D002118:Calcium