Regulation of the sarcoplasmic reticulum Ca(2+)-release channel requires intact annexin VI.
Annexin VI has eight highly conserved repeated domains; all other annexins have four. Díaz-Muñoz et al. (J Biol Chem 265:15894, 1990) reported that annexin VI alters the gating properties of the ryanodine-sensitive Ca(2+)-release channel isolated from sarcoplasmic reticulum. The investigate the domain structure of rat annexin VI (67 kDa calcimedin) required for this channel regulation, various proteolytic digestions were performed. In each case, protease-resistant core polypeptides were produced. Annexin VI was digested with V8 protease and two core polypeptides were purified by Ca(2+)-dependent phospholipid binding followed by HPLC. The purified fragments were shown to be derived from the N- and C-terminal halves of annexin VI, and demonstrated differential immunoreactivity with monoclonal antibodies to rat annexin VI. While both core polypeptides retained their ability to bind phospholipids in a Ca(2+)-dependent manner, they did not regulate the sarcoplasmic reticulum Ca(2+)-dependent manner, they did not regulate the sarcoplasmic reticulum Ca(2+)-release channel as did intact annexin VI.
Hazarika
P
P
Department of Physiology and Cell Biology, University of Texas Medical School, Houston 77225.
Sheldon
A
A
Kaetzel
M A
MA
Díaz-Muñoz
M
M
Hamilton
S L
SL
Dedman
J R
JR
D000595:Amino Acid Sequence; D000818:Animals; D017317:Annexin A6; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000502:physiology*; D002918:Chymotrypsin / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D008051:Lipid Bilayers; D008969:Molecular Sequence Data; D010455:Peptides / Q000502:physiology; D011402:Pronase / Q000378:metabolism; D051381:Rats; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D017317:Annexin A6; D015220:Calcium Channels; D002135:Calcium-Binding Proteins; D008051:Lipid Bilayers; D010455:Peptides; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C028344:glutamyl endopeptidase; D014357:Trypsin; D011402:Pronase; D002118:Calcium