Measurement of Resting Cytosolic Ca2+ Concentrations and Ca2+ Store Size in HEK-293 Cells Transfected with Malignant Hyperthermia or Central Core Disease Mutant Ca2+ Release Channels*
Measurement of Resting Cytosolic Ca2+ Concentrations and Ca2+ Store Size in HEK-293 Cells Transfected with Malignant Hyperthermia or Central Core Disease Mutant Ca2+ Release Channels*
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测量转染恶性高热或中枢核心疾病突变 Ca2 释放通道的 HEK-293 细胞中静息胞质 Ca2 浓度和 Ca2 储存大小*
DOI：
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发表时间：
1999
影响因子：
4.8
通讯作者：
D. Maclennan
中科院分区：
文献类型：
--
作者：
Jiefei Tong;T. McCarthy;D. Maclennan
将恶性高热（MH）和中央核心病（CCD）突变引入全长兔钙释放通道（RYR 1）cDNA，然后在HEK-293细胞中瞬时表达。静息Ca 2+浓度在表达同源四聚体CCD突变体RyR 1的HEK-293细胞中高于表达同源四聚体MH突变体RyR 1的细胞。表达同源四聚体CCD或MH突变体RyR 1的细胞表现出较低的咖啡因诱导的Ca 2+释放的最大峰值振幅比表达野生型RyR 1的细胞，这表明MH和CCD突变体可能是“泄漏”。在表达同源四聚体野生型或突变体RyR 1的细胞中，10 mm咖啡因诱导的Ca 2+释放的幅度与卡巴胆碱或毒胡萝卜素诱导的Ca 2+释放的幅度显著相关，表明最大药物诱导的Ca 2+释放取决于内质网Ca 2+储存的大小。通过酶联免疫吸附试验、45 Ca 2+摄取和共聚焦显微镜测定，表达野生型或突变型RyR 1的HEK-293细胞中内源性肌浆网（内）Ca 2 +-ATP酶亚型2b（SERCA 2b）的含量增加，支持内质网Ca 2+储存能力增加作为对增强的Ca 2+泄漏的补偿反应的观点。当MH/CCD突变体和野生型RyR 1的异源四聚体（1：1）组合与SERCA 1一起表达以增强Ca 2+再摄取时，响应于低浓度咖啡因和氟烷的Ca 2+释放的幅度高于在表达野生型RyR 1和SERCA 1的细胞中观察到的幅度。在无钙培养基中，MH/CCD突变体比野生型RyR 1对咖啡因更敏感，表明用各种MH/CCD突变体RyR 1蛋白观察到的咖啡因超敏反应不依赖于细胞外Ca 2+浓度。
Malignant hyperthermia (MH) and central core disease (CCD) mutations were introduced into full-length rabbit Ca2+ release channel (RYR1) cDNA, which was then expressed transiently in HEK-293 cells. Resting Ca2+ concentrations were higher in HEK-293 cells expressing homotetrameric CCD mutant RyR1 than in cells expressing homotetrameric MH mutant RyR1. Cells expressing homotetrameric CCD or MH mutant RyR1 exhibited lower maximal peak amplitudes of caffeine-induced Ca2+ release than cells expressing wild type RyR1, suggesting that MH and CCD mutants might be “leaky.” In cells expressing homotetrameric wild type or mutant RyR1, the amplitude of 10 mm caffeine-induced Ca2+ release was correlated significantly with the amplitude of carbachol- or thapsigargin-induced Ca2+ release, indicating that maximal drug-induced Ca2+ release depends on the size of the endoplasmic reticulum Ca2+ store. The content of endogenous sarco(endo)plasmic reticulum Ca2+-ATPase isoform 2b (SERCA2b), measured by enzyme-linked immunosorbent assay,45Ca2+ uptake, and confocal microscopy, was increased in HEK-293 cells expressing wild type or mutant RyR1, supporting the view that endoplasmic reticulum Ca2+ storage capacity is increased as a compensatory response to an enhanced Ca2+ leak. When heterotetrameric (1:1) combinations of MH/CCD mutant and wild type RyR1 were expressed together with SERCA1 to enhance Ca2+ reuptake, the amplitude of Ca2+release in response to low concentrations of caffeine and halothane was higher than that observed in cells expressing wild type RyR1 and SERCA1. In Ca2+-free medium, MH/CCD mutants were more sensitive to caffeine than wild type RyR1, indicating that caffeine hypersensitivity observed with a variety of MH/CCD mutant RyR1 proteins is not dependent on extracellular Ca2+ concentration.
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成熟禽类快肌骨骼肌中两种三联体连接足蛋白亚型的鉴定和定位。
DOI：
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发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Airey,JA;Beck,CF;Murakami,K;Tanksley,SJ;Deerinck,TJ;Ellisman,MH;Sutko,JL
通讯作者：
Sutko,JL
SR在恶性高热中的作用。
影响因子：
4
作者：
Nelson,TE
通讯作者：
Nelson,TE
影响因子：
56.9
作者：
MA, J;FILL, M;CORONADO, R
通讯作者：
CORONADO, R
猪恶性高热易感性：骨骼肌肌浆网超敏钙释放机制。
DOI：
--
发表时间：
1986
期刊：
Canadian journal of veterinary research = Revue canadienne de recherche veterinaire
影响因子：
--
作者：
O'Brien,PJ
通讯作者：
O'Brien,PJ
影响因子：
18.2
作者：
Martonosi,A
通讯作者：
Martonosi,A