Isolation of a protein fraction from Bordetella pertussis that facilitates entry of the calmodulin-sensitive adenylate cyclase into animal cells.
Isolation of a protein fraction from Bordetella pertussis that facilitates entry of the calmodulin-sensitive adenylate cyclase into animal cells.
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从百日咳博德特氏菌中分离出蛋白质组分，促进钙调素敏感的腺苷酸环化酶进入动物细胞。
DOI：
10.1021/bi00446a024
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发表时间：
1989
期刊：
影响因子：
2.9
通讯作者：
Storm,DR
中科院分区：
文献类型：
--
作者：
Donovan,MG;Masure,HR;Storm,DR
药理学系，SJ-30，华盛顿大学医学院，西雅图，华盛顿 98195 收稿日期：1989 年 4 月 14 日；修订稿于 1989 年 6 月 13 日收到摘要：百日咳博德特氏菌（Bordetella pertussis）是引起百日咳的病原体，它向其培养基中释放一种可溶性钙调蛋白敏感的腺苷酸环化酶。一些研究人员表明，部分纯化的腺苷酸环化酶能够进入动物细胞并提高细胞内 cAMP 水平 [Confer, D. L., & Eaton, J. W.(1982) Science 217, 948-950； Shattuck, RL, & Storm, DR (1985) Biochemistry 24, 6323-6328]，然而，腺苷酸环化酶催化亚基进入动物细胞的机制尚不清楚。最近，已确定该酶的纯化催化亚基无法进入动物细胞[Masure, H. R., Oldenburg, D. J., Donovan, M. G., Shattuck, R. L., & Storm, DR (1988) J. Biol.化学。 263、6933-6940]。基于这些数据和其他观察结果，我们假设百日咳博德特氏菌的培养基含有一种或多种促进腺苷酸环化酶催化亚基进入动物细胞的附加多肽。在这项研究中，我们报告了百日咳博德特氏菌腺苷酸环化酶的细胞侵入性制剂在通过麦芽凝集素-琼脂糖柱后变得非侵入性。用N-乙酰基-D-葡糖胺从麦芽凝集素-琼脂糖柱洗脱级分。当该部分与非侵入性腺苷酸环化酶结合时，能够恢复腺苷酸环化酶制剂对肠神经母细胞瘤细胞的能力并增加细胞内cAMP水平。此外，发现从麦芽凝集素-琼脂糖柱洗脱的级分对胰蛋白酶和糜蛋白酶敏感，表明该材料是蛋白质。麦芽凝集素-琼脂糖柱洗脱液的 SDS 凝胶电泳显示存在三种表观分子量值为 26、28 和 30 kDa 的多肽。这些数据提供了第一个直接证据，证明百日咳博德特氏菌产生的额外的、可分离的蛋白质成分是催化亚基进入动物细胞所必需的。百日咳博德特氏菌是一种小型革兰氏阴性杆菌，是百日咳的病原体（Olson，1975；Wardlaw & Parton，1988）。百日咳博德特氏菌生长的培养基含有许多生物活性成分，这些成分被认为在该疾病的发病机制中发挥作用。其中之一，胰岛激活蛋白 (IAP)，1 已被纯化并显示可减弱多种哺乳动物细胞中受体介导的腺苷酸环化酶抑制
Department of Pharmacology, SJ-30, School of Medicine, University of Washington, Seattle, Washington 98195 Received April 14, 1989; Revised Manuscript Received June 13, 1989 abstract: Bordetella pertussis, the pathogen responsible for whoopingcough, releases a soluble calmodulin-sensitive adenylate cyclase into its culture medium. Several investigators have shown that the partially purified adenylate cyclase is capable of entering animal cells and elevating intracellular cAMP levels [Confer, D. L., & Eaton, J. W.(1982) Science 217, 948-950; Shattuck, RL, & Storm, DR (1985) Biochemistry 24, 6323-6328], However, the mechanism for entry of the catalytic subunit of the adenylatecyclase into animal cells is unknown. Recently, it was determined that the purified catalytic subunit of the enzyme is unable to enter animal cells [Masure, H. R., Oldenburg, D. J., Donovan, M. G., Shattuck, R. L., & Storm, DR (1988) J. Biol. Chem. 263, 6933-6940]. Onthe basis of these data and other observations, we hypothesized that the culture medium of B. pertussis contains one or more additional polypeptides which facilitate entry of the adenylate cyclase catalytic subunit into animal cells. In this study, we report that a cell-invasive preparation of B. pertussis adenylate cyclase was rendered noninvasive after passage through a wheat germ lectin-agarose column. A fraction was eluted from the wheat germ lectin-agarose column with iV-acetyl-D-glucosamine. This fraction, when combined with the noninvasive adenylate cyclase, was able to restore the ability of the adenylate cyclase preparation to enterneuroblastoma cells and increase intracellular cAMP levels. Furthermore, the fraction eluted from the wheat germ lectin-agarose column was found to be trypsin and chymotrypsin sensitive, suggesting that this material was proteinaceous. SDS gel electrophoresis of the eluate from the wheat germ lectin-agarose column revealed the presence of three polypeptides with apparent molecular mass values of 26, 28, and 30 kDa. These data provide the first direct evidence for the existence of an additional, separable protein component produced by B. pertussisthat is required for entry of the catalytic subunit into animal cells.Bordetella pertussis is a small, Gram-negative bacillus that is the pathogen responsible for whoopingcough (Olson, 1975; Wardlaw & Parton, 1988). The culture medium of growing B. pertussis contains a number of biologically active components which are thought to play a role in the pathogenesis of the disease. One of these, islet activating protein (IAP), 1 has been purified and shown to attenuate receptor-mediated in-hibition of adenylate cyclase in a variety of mammalian cell
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DOI：
10.1099/00221287-63-2-211
发表时间：
1970-01-01
期刊：
JOURNAL OF GENERAL MICROBIOLOGY
影响因子：
--
作者：
STAINER, DW;SCHOLTE, MJ
通讯作者：
SCHOLTE, MJ
百日咳博德特氏菌的侵入性腺苷酸环化酶。
DOI：
10.1042/bj2430145
发表时间：
1987
期刊：
The Biochemical journal
影响因子：
--
作者：
Eitan Friedman;Zvi FARFELt;Emanuel Hanski
通讯作者：
Emanuel Hanski
百日咳博德特氏菌钙调素敏感腺苷酸环化酶：在大肠杆菌中的克隆和表达
影响因子：
3.6
作者：
P. Glaser;D. Ladant;O. Sezer;F. Pichot;A. Ullmann;A. Danchin
通讯作者：
A. Danchin
DOI：
10.1016/s0021-9258(18)66710-9
发表时间：
1986-12
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Daniel Ladant;Colette Brezins;Jean-Michel Alonsos;Isabelle CrenonS;Nicole Guiso
通讯作者：
Daniel Ladant;Colette Brezins;Jean-Michel Alonsos;Isabelle CrenonS;Nicole Guiso
钙调蛋白对腺苷酸环化酶的非钙依赖性激活。
DOI：
10.1111/j.1432-1033.1983.tb07423.x
发表时间：
1983
期刊：
European journal of biochemistry
影响因子：
--
作者：
M. Kilhoffer;G. H. Cook;J. Wolff
通讯作者：
J. Wolff