STIMULATION OF PHOSPHOLIPASE C-BETA(2) BY RECOMBINANT GUANINE-NUCLEOTIDE-BINDING PROTEIN BETA-GAMMA DIMERS PRODUCED IN A BACULOVIRUS/INSECT CELL EXPRESSION SYSTEM - REQUIREMENT OF GAMMA-SUBUNIT ISOPRENYLATION FOR STIMULATION OF PHOSPHOLIPASE-C
STIMULATION OF PHOSPHOLIPASE C-BETA(2) BY RECOMBINANT GUANINE-NUCLEOTIDE-BINDING PROTEIN BETA-GAMMA DIMERS PRODUCED IN A BACULOVIRUS/INSECT CELL EXPRESSION SYSTEM - REQUIREMENT OF GAMMA-SUBUNIT ISOPRENYLATION FOR STIMULATION OF PHOSPHOLIPASE-C
复制标题
DOI：
10.1111/j.1432-1033.1994.tb19927.x
复制
发表时间：
1994-01-15
期刊：
影响因子：
--
通讯作者：
GIERSCHIK, P
中科院分区：
文献类型：
--
作者：
DIETRICH, A;MEISTER, M;GIERSCHIK, P
在杆状病毒感染的昆虫细胞中表达了重组野生型β (1) γ(1)二聚体信号转导鸟嘌呤核苷酸结合蛋白（G蛋白）和β (1) γ(1)二聚体携带已知阻断γ -亚基异戊二烯化的突变(β (1) γ (1)C71S)。野生型和突变型β (1) γ(1)二聚体均可在感染细胞的可溶性部分亚细胞分离中发现。阴离子交换色谱和代谢放射性标记研究表明，可溶性β (1) γ(1)制剂含有大约等量的非异丙烯化和异丙烯化β (1) γ(1)二聚体。用重组磷脂酶c - β(2)重组了从牛视网膜中纯化的可溶性野生型和突变型β (1) γ(1)二聚体和天然β (1) γ(1)二聚体。只有异戊烯基化的β (1) γ(1)二聚体能够刺激磷脂酶c - β(2)。结果表明，β - γ亚基异戊烯酰化和/或额外的蛋白质翻译后加工是β - γ亚基刺激磷脂酶C所必需的。
Recombinant wild-type beta(1) gamma(1) dimers of signal-transducing guanine nucleotide-binding proteins (G proteins) and beta(1) gamma 1 dimers carrying a mutation known to block gamma-subunit isoprenylation (beta(1) gamma(1)C71S) were expressed in baculovirus-infected insect cells. Both wild-type and mutant beta(1) gamma(1) dimers were found in soluble fractions of infected cells upon subcellular fractionation. Anion exchange chromatographic and metabolic-radiolabeling studies revealed that the soluble beta(1) gamma(1) preparation contained approximately equal amounts of non-isoprenylated and isoprenylated beta(1) gamma(1) dimers. Soluble wild-type and mutant beta(1) gamma(1) dimers and native beta(1) gamma(1) dimers purified from bovine retina were reconstituted with recombinant phospholipase C-beta(2). Only isoprenylated beta(1) gamma(1) dimers were capable of stimulating phospholipase C-beta(2). The results show that gamma-subunit isoprenylation and/or additional post-translational processing of the protein are required for beta gamma subunit stimulation of phospholipase C.