Abstract
Transverse tubule (TT) membrane vesicles have been isolated from the skeletal muscle of normal and malignant hyperthermia-susceptible (MHS) pigs. MHS and normal TT did not differ in the distribution of the major proteins, cholesterol, or phospholipid content, (Na+ + K+)-ATPase activity, [3H]ouabain binding, Ca2+-ATPase activity, Mg2+-ATPase activity, or [3H]saxitoxin binding. Furthermore, in the presence of micromolar Ca2+, MHS and normal TT did not differ significantly in the K(D) values for either [3H]nitrendipine binding (2.7 ± 0.6 and 3.3 ± 0.5 nM, respectively) or (-)-[3H]desmethoxyverapamil ([3H]D888) binding (7.2 ± 0.9 and 6.4 ± 0.6 nM, respectively). However, in contrast to normal TT, MHS TT exhibited a significantly decreased B(max) for both [3H]nitrendipine binding (26.4 ± 5.4 for MHS versus 40.6 ± 3.7 pmol/mg protein for normal TT) and [3H]D888 binding (17.8 ± 7.0 for MHS versus 37.4 ± 5.9 pmol/mg protein for normal TT). At calcium concentrations greater than 0.1 mM, there was a greater inhibition of [3H]nitrendipine binding to normal than to MHS TT such that binding was now similar for both preparations. As with purified TT, [3H]nitrendipine binding to MHS muscle homogenates was significantly less than to normal muscle homogenates (109 ± 20 versus 211 ± 19 pmol/mg protein, for MHS and normal TT, respectively); this difference was not apparent when 100 mM CaCl2 was included in the binding medium. We conclude that the altered MHS TT dihydropyridine receptor properties may reflect an adaptation of the TT voltage sensing mechanism to the abnormal sarcoplasmic reticulum calcium release channel regulation in MHS muscle.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2711-2717 |
| Number of pages | 7 |
| Journal | Journal of Biological Chemistry |
| Volume | 264 |
| Issue number | 5 |
| State | Published - Jan 1 1989 |
Fingerprint
Cite this
Altered transverse tubule dihydropyridine receptor binding in malignant hyperthermia. / Ervasti, James M; Claessens, M. T.; Mickelson, James R; Louis, C. F.
In: Journal of Biological Chemistry, Vol. 264, No. 5, 01.01.1989, p. 2711-2717.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Altered transverse tubule dihydropyridine receptor binding in malignant hyperthermia
AU - Ervasti, James M
AU - Claessens, M. T.
AU - Mickelson, James R
AU - Louis, C. F.
PY - 1989/1/1
Y1 - 1989/1/1
N2 - Transverse tubule (TT) membrane vesicles have been isolated from the skeletal muscle of normal and malignant hyperthermia-susceptible (MHS) pigs. MHS and normal TT did not differ in the distribution of the major proteins, cholesterol, or phospholipid content, (Na+ + K+)-ATPase activity, [3H]ouabain binding, Ca2+-ATPase activity, Mg2+-ATPase activity, or [3H]saxitoxin binding. Furthermore, in the presence of micromolar Ca2+, MHS and normal TT did not differ significantly in the K(D) values for either [3H]nitrendipine binding (2.7 ± 0.6 and 3.3 ± 0.5 nM, respectively) or (-)-[3H]desmethoxyverapamil ([3H]D888) binding (7.2 ± 0.9 and 6.4 ± 0.6 nM, respectively). However, in contrast to normal TT, MHS TT exhibited a significantly decreased B(max) for both [3H]nitrendipine binding (26.4 ± 5.4 for MHS versus 40.6 ± 3.7 pmol/mg protein for normal TT) and [3H]D888 binding (17.8 ± 7.0 for MHS versus 37.4 ± 5.9 pmol/mg protein for normal TT). At calcium concentrations greater than 0.1 mM, there was a greater inhibition of [3H]nitrendipine binding to normal than to MHS TT such that binding was now similar for both preparations. As with purified TT, [3H]nitrendipine binding to MHS muscle homogenates was significantly less than to normal muscle homogenates (109 ± 20 versus 211 ± 19 pmol/mg protein, for MHS and normal TT, respectively); this difference was not apparent when 100 mM CaCl2 was included in the binding medium. We conclude that the altered MHS TT dihydropyridine receptor properties may reflect an adaptation of the TT voltage sensing mechanism to the abnormal sarcoplasmic reticulum calcium release channel regulation in MHS muscle.
AB - Transverse tubule (TT) membrane vesicles have been isolated from the skeletal muscle of normal and malignant hyperthermia-susceptible (MHS) pigs. MHS and normal TT did not differ in the distribution of the major proteins, cholesterol, or phospholipid content, (Na+ + K+)-ATPase activity, [3H]ouabain binding, Ca2+-ATPase activity, Mg2+-ATPase activity, or [3H]saxitoxin binding. Furthermore, in the presence of micromolar Ca2+, MHS and normal TT did not differ significantly in the K(D) values for either [3H]nitrendipine binding (2.7 ± 0.6 and 3.3 ± 0.5 nM, respectively) or (-)-[3H]desmethoxyverapamil ([3H]D888) binding (7.2 ± 0.9 and 6.4 ± 0.6 nM, respectively). However, in contrast to normal TT, MHS TT exhibited a significantly decreased B(max) for both [3H]nitrendipine binding (26.4 ± 5.4 for MHS versus 40.6 ± 3.7 pmol/mg protein for normal TT) and [3H]D888 binding (17.8 ± 7.0 for MHS versus 37.4 ± 5.9 pmol/mg protein for normal TT). At calcium concentrations greater than 0.1 mM, there was a greater inhibition of [3H]nitrendipine binding to normal than to MHS TT such that binding was now similar for both preparations. As with purified TT, [3H]nitrendipine binding to MHS muscle homogenates was significantly less than to normal muscle homogenates (109 ± 20 versus 211 ± 19 pmol/mg protein, for MHS and normal TT, respectively); this difference was not apparent when 100 mM CaCl2 was included in the binding medium. We conclude that the altered MHS TT dihydropyridine receptor properties may reflect an adaptation of the TT voltage sensing mechanism to the abnormal sarcoplasmic reticulum calcium release channel regulation in MHS muscle.
UR - http://www.scopus.com/inward/record.url?scp=0024537210&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0024537210&partnerID=8YFLogxK
M3 - Article
C2 - 2536721
AN - SCOPUS:0024537210
VL - 264
SP - 2711
EP - 2717
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 5
ER -