PROTEIN-PROTEIN INTERACTIONS IN THE 18S ATPASE OF CHLAMYDOMONAS OUTER DYNEIN ARMS
PROTEIN-PROTEIN INTERACTIONS IN THE 18S ATPASE OF CHLAMYDOMONAS OUTER DYNEIN ARMS
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DOI：
10.1002/cm.970060510
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发表时间：
1986-01-01
影响因子：
--
通讯作者：
ROSENBAUM, JL
中科院分区：
文献类型：
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作者：
MITCHELL, DR;ROSENBAUM, JL
当从衣原体鞭毛轴丝中提取外排动力蛋白臂时，它们解离成两个ATP酶复合物，其沉降系数为12 S和18 S。我们用18 S动力蛋白免疫小鼠，并产生了针对该复合物中多肽的单克隆抗体库。选择特异性识别18 S α-和β-重链以及83，000-道尔顿和70，000-道尔顿中间链。这些抗体被分离和表征其识别变性抗原和天然18 S动力蛋白上的决定簇的能力; 18 S动力蛋白以逐步的方式用离子和非离子去污剂解离成较小的聚集体，并且通过用特异性单克隆抗体沉淀分离所得的亚复合物。分离的最小聚集体是α-β-葡聚糖之间的异二聚体。链和16，000-道尔顿轻链以及两个中间链之间。另外的β-还观察到具有18，000-道尔顿轻链和70，000-道尔顿中间链的重链，以及中间链异二聚体与21，000道尔顿和12，500道尔顿轻链之间的较弱相互作用。我们提出了一个模型的18 S动力蛋白亚结构的基础上，这些信息。
When outer-row dynein arms are extracted from Chlamydomonas flagellar axonemes, they dissociate into two ATPase complexes with sedimentation coefficients of 12S and 18S. We immunized mice with 18S dynein and generated a library of monoclonal antibodies against the polypeptides in this complex. Antibodies were selected which specifically recognize the 18S .alpha.- and .beta.-heavy chains and the 83,000-dalton and 70,000-dalton intermediate chains. These antibodies were isolated and characterized for their ability to recognize determinants on both denatured antigens and native 18S dynein; 18S dynein was dissociated in stepwise fashion into smaller aggregates with ionic and nonionic detergents and the resulting subcomplexes were isolated by precipitation with specific monoclonal antibodies. The smallest aggregates isolated were heterodimers between the .alpha.-chain and a 16,000-dalton light chain and between the two intermediate chains. Additional close associations of the .beta.-heavy chain with an 18,000-dalton light chain and 70,000-dalton intermediate chain, and a weaker interaction between the intermediate chain heterodimer and light chains of 21,000 daltons and 12,500 daltons, were also observed. We present a model of 18S dynein substructure based upon this information.