Involvement of protein serine and threonine phosphorylation in human sperm capacitation
Involvement of protein serine and threonine phosphorylation in human sperm capacitation
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DOI：
10.1095/biolreprod60.6.1402
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发表时间：
1999-06-01
影响因子：
3.6
通讯作者：
Naz, RK
中科院分区：
文献类型：
--
作者：
Naz, RK
研究了丝氨酸和苏氨酸磷酸化在人类精子获能中的作用。抗磷酸丝氨酸单抗在43-55 kDa、94+/-2 kDa、110 kDa和190 kDa分子区段识别6条蛋白质条带，在18 kDa和35 kDa区域各识别一条微弱条带。除18 kDa、35 kDa和94+/-2 kDa蛋白条带更清晰、更粗，在110 kDa分子区域还观察到一条额外的条带外，抗磷酸苏氨酸单抗在6个相似区域识别蛋白质条带。在43-55 kDa的分子区域，有一种特性良好的糖蛋白，命名为受精抗原，它显示在溶解的人透明带暴露后，丝氨酸/苏氨酸磷酸化进一步增加。在微球或溶液中进行的体外无细胞激酶分析中，属于相似分子区域的4~8个蛋白质，即20+/-2 kDa、43-55 kDa、94+/-2 kDa和110+/-10 kDa，以及80+/-4和210+/-10 kDa，在双残基(丝氨酸/酪氨酸和苏氨酸/酪氨酸)上被磷酸化。获能增加了每个精子细胞的丝氨酸/苏氨酸磷酸化强度，增加了被磷酸化的精子细胞的数量，并诱导了丝氨酸/苏氨酸特异性荧光的亚细胞位移。这些发现表明，蛋白质丝氨酸/苏氨酸磷酸化参与了精子获能，并可能在精子获能中起到生理作用。
The involvement of serine and threonine phosphorylation in human sperm capacitation was investigated. Anti-phosphoserine monoclonal antibody (mAb) recognized six protein bands in the 43-55-kDa, 94 +/- 2-kDa, 110-kDa, and 190-kDa molecular regions, in addition to a faint band each in the 18-kDa and 35-kDa regions. Anti-phosphothreonine mAb recognized protein bands in six similar regions, except that the 18-kDa, 35-kDa, and 94 +/- 2-kDa protein bands were sharper and thicker, and an additional band was observed in the 110-kDa molecular region. In the 43-55-kDa molecular region, there was a well-characterized glycoprotein, designated fertilization antigen, that showed a further increase in serine/threonine phosphorylation after exposure to solubilized human zona pellucida. In a cell-free in vitro kinase assay carried out on beads or in solution, four to eight proteins belonging to similar molecular regions, namely 20 +/- 2 kDa, 43-55 kDa, 94 +/- 2 kDa, and 110 +/- 10 kDa, as well as in 80 +/- 4 and 210 +/- 10 kDa regions, were phosphorylated at dual residues (serine/tyrosine and threonine/tyrosine). Capacitation increased the intensity of serine/threonine phosphorylation per sperm cell, increased the number of sperm cells that were phosphorylated, and induced a subcellular shift in the serine/threonine-specific fluorescence. These findings indicate that protein serine/threonine phosphorylation is involved and may have a physiological role in sperm capacitation.