Jin Z X, Inaba K, Manaka K, Morisawa M, Hayashi H
Sugashima Marine Biological Laboratory, School of Science, Nagoya University, Mie.
J Biochem. 1994 May;115(5):885-90. doi: 10.1093/oxfordjournals.jbchem.a124435.
We isolated and characterized several monoclonal antibodies against a protein complex containing the flagellar movement-initiating phosphoprotein (MIPP) that appears to play a crucial role in the initiation of flagellar movement in quiescent spermatozoa of Salmonid fish. The effects of the antibodies on the phosphorylation of MIPP, as well as on the initiation of movement, in model sperm cells were studied. Three monoclonal antibodies, namely, FMI7, FMI18, and FMI27, were found specifically to inhibit both the initiation of flagellar movement and the phosphorylation of MIPP. These antibodies did not recognize denatured MIPP; they only recognized the native antigen. FMI7 exclusively recognized the denatured form of a 38-kDa protein, which may possibly be a protein kinase responsible for the phosphorylation of MIPP. Immunofluorescence analysis in situ of model sperm cells with the antibodies showed that the antigen was localized predominantly in the basal structure of the spermatozoon. Thus, the results clearly demonstrate the involvement of MIPP in the initiation of flagellar movement and the control of flagellar motility.
我们分离并鉴定了几种针对一种蛋白质复合物的单克隆抗体，该蛋白质复合物包含鞭毛运动起始磷蛋白（MIPP），而MIPP似乎在鲑科鱼类静止精子的鞭毛运动起始过程中起着关键作用。我们研究了这些抗体对模型精子细胞中MIPP磷酸化以及运动起始的影响。发现三种单克隆抗体，即FMI7、FMI18和FMI27，能特异性抑制鞭毛运动的起始和MIPP的磷酸化。这些抗体不能识别变性的MIPP；它们只识别天然抗原。FMI7专门识别一种38 kDa蛋白质的变性形式，该蛋白质可能是负责MIPP磷酸化的蛋白激酶。用这些抗体对模型精子细胞进行原位免疫荧光分析表明，抗原主要定位于精子的基部结构。因此，结果清楚地证明了MIPP参与鞭毛运动的起始和鞭毛运动的控制。
