GTP BINDING PROTEINS AND ADENYLATE CYCLASE
GTP 结合蛋白和腺苷酸环化酶
基本信息
- 批准号:3843261
- 负责人:
- 金额:--
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:
- 资助国家:美国
- 起止时间:至
- 项目状态:未结题
- 来源:
- 关键词:ADP ribosylation RNA splicing Saccharomyces cerevisiae adenylate cyclase cholera toxin cow density gradient ultracentrifugation enzyme activity enzyme substrate gene expression genetic promoter element genetic transcription guanine nucleotide binding protein laboratory rabbit laboratory rat molecular cloning western blottings
项目摘要
Six mammalian ADP-ribosylation factors (ARFs) identified by cDNA cloning
were expressed as recombinant proteins (rARF) that stimulated cholera
toxin ADP-ribosyltransferase activity. Microsequencing of sARF I and
II, purified from bovine brain (Tsai et al. (1988) J. Biol. Chem. 263:
1768-1772), established that they are ARF 1 and 3, respectively. Rabbit
antibodies (IgG) against sARF II reacted similarly with ARFs 1, 2, and 3
(class I) on Western blots. ARF 1 and 3 were distinquished by their
electrophoretic mobilities; ARF 2 was not detected in rat brain. Rabbit
antiserum against rARF 5 cross-reacted partially with rARF 4, but not
detectably with rARF 6 and minimally with class I ARFs. ARF 4, which
differs from ARF 5 in electrophoretic mobility, was not detected in rat
brain. GTPgammaS increased recovery of ARF activity and
immunoreactivity in organelle fractions separated by density gradient
centrifugation, after incubation of rat brain homogenate with ATP and a
regenerating system.
Saccharomyces cerevisiae has two ARF proteins that are 96% identical.
The ARF 1 gene is constitutively expressed whereas the ARF 2 gene is
repressed by glucose. Expression of human ARF 5 or ARF 6 or Giardia ARF
can resume much more slowly, however, than wild-type yeast or mutants
rescued with yeast ARF 1, from which we infer that the heterologous ARFs
may not function efficiently or effectively in the yeast vesicular
protein trafficking system.
哺乳动物ADP核糖基化因子(ARF)的cDNA克隆
表达为重组蛋白(rARF),
毒素ADP-核糖基转移酶活性。 sARF I和
II,从牛脑中纯化(Tsai等(1988)J. Biol. Chem. 263:
1768 - 1772),确定它们分别是ARF 1和3。 兔
抗sARF II的抗体(IgG)与ARF 1、2和3的反应相似
(I类)蛋白质印迹法。 ARF 1和3的区别在于它们的
电泳迁移率; ARF 2在大鼠脑中未检测到。 兔
抗rARF 5的抗血清与rARF 4部分交叉反应,
rARF 6可检测,I类ARF最低。 ARF 4,
与ARF 5的电泳迁移率不同,在大鼠中未检测到
个脑袋 GTP γ S增加ARF活性的恢复,
通过密度梯度分离的细胞器组分中的免疫反应性
离心后,大鼠脑匀浆与ATP和a
再生系统
酿酒酵母有两种ARF蛋白,它们96%相同。
ARF 1基因是组成型表达的,而ARF 2基因是组成型表达的。
被葡萄糖抑制 人ARF 5或ARF 6或贾第虫ARF的表达
但是,与野生型酵母或突变体相比,
用酵母ARF 1拯救,从中我们推断异源ARF
在酵母囊泡中可能不能有效地发挥作用,
蛋白质运输系统
项目成果
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专利数量(0)
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