Modulation of Ca2+ release channel activity from sarcoplasmic reticulum by annexin VI (67-kDa calcimedin).
The effect of annexin VI (67-kDa calcimedin) on the activity of the Ca2+ release channel was studied using heavy sarcoplasmic reticulum membranes reconstituted into planar bilayers. Annexin VI, in a range of 5-40 nM, modified the gating behavior of the Ca2+ release channel by increasing the probability of opening by 2.7-fold and the mean open time by 82-fold relative to controls. Annexin VI caused no change in the slope conductance of the channel. The modulatory effect of annexin VI on the activity of Ca2+ release channels was Ca2+ dependent, and the annexin VI-modified channel was sensitive to both ruthenium red and ryanodine. The effect of annexin VI was observed when this protein was added specifically to the trans chamber, which corresponds to the luminal side of sarcoplasmic reticulum as determined by the ATP activation of the channel. In addition, differential extraction studies demonstrated that some annexin VI is localized within the lumen of the isolated heavy sarcoplasmic reticulum vesicles prepared by several different procedures. Annexin VI did not modify, from either the cis or trans chambers, the activity of K+ or Cl- channels from sarcoplasmic reticulum or the dihydropyridine sensitive Ca2+ channel from transverse tubules. In addition, the 38-kDa core proteolytic fragments of annexin VI had no effect on the Ca2+ release channel activity. Annexin VI is therefore a candidate for a physiological modulator of the Ca2+ release channel and as such, may play an important role in the excitation-contraction coupling.
Díaz-Muñoz
M
M
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas 77030.
Hamilton
S L
SL
Kaetzel
M A
MA
Hazarika
P
P
Dedman
J R
JR
D000818:Animals; D017317:Annexin A6; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D015220:Calcium Channels / Q000187:drug effects / Q000502:physiology*; D002135:Calcium-Binding Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D008051:Lipid Bilayers; D008564:Membrane Potentials / Q000187:drug effects; D008861:Microsomes / Q000502:physiology; D009132:Muscles / Q000502:physiology; D011817:Rabbits; D051381:Rats; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000502:physiology*; D013552:Swine
D017317:Annexin A6; D015220:Calcium Channels; D002135:Calcium-Binding Proteins; D008051:Lipid Bilayers; D002118:Calcium