Tash J S, Bracho G E
Department of Molecular and Integrative Physiology and the Center for Reproductive Sciences, University of Kansas Medical Center, Kansas City, Kansas, 66160, USA.
Biochem Biophys Res Commun. 1998 Oct 20;251(2):557-63. doi: 10.1006/bbrc.1998.9516.
A method for collecting live immotile cauda epididymal mouse sperm that initiate motility by dilution into an activation buffer is described. Sperm in collection buffer showed low percent motility (MOT) and population progression (PRG) that increased 10-fold and 9-fold, respectively, during the first 2 min after dilution into activation buffer. Western phosphoserine (pS), phosphothreonine (pT), and phosphotyrosine (pY) analysis revealed a 120 kDa protein that markedly increased in pT content during initiation of motility and may be related to FP130, the motility-coupled axonemal protein of sea urchin sperm. A prominent 82 kDa protein that was pS and pT-phosphorylated in immotile and motile sperm is likely the fibrous sheath component AKAP82 that is phosphorylated during spermatogenesis. Analysis of live human sperm also identified a prominent 120 kDa pT protein. Thus it appears that phosphorylation of FP130 and related 120 kDa proteins in mouse, and perhaps human sperm, represent common targets during motility initiation in sperm.
描述了一种收集活的、无运动能力的附睾尾小鼠精子的方法，这些精子通过稀释到激活缓冲液中而开始运动。收集缓冲液中的精子显示出低运动百分比（MOT）和群体前进率（PRG），在稀释到激活缓冲液后的最初2分钟内，它们分别增加了10倍和9倍。蛋白质免疫印迹法检测磷酸化丝氨酸（pS）、磷酸化苏氨酸（pT）和磷酸化酪氨酸（pY）发现，有一种120 kDa的蛋白质在精子开始运动时pT含量显著增加，可能与海胆精子的运动偶联轴丝蛋白FP130有关。一种在无运动能力和有运动能力的精子中均被pS和pT磷酸化的82 kDa的显著蛋白质，可能是在精子发生过程中被磷酸化的纤维鞘成分AKAP82。对活人精子的分析也鉴定出一种显著的120 kDa的pT蛋白质。因此，小鼠精子中FP130和相关的120 kDa蛋白质的磷酸化，可能还有人类精子中的，似乎代表了精子运动起始过程中的共同靶点。
