PHOSPHORYLATION OF HUMAN SPERM PROTAMINES HP1 AND HP2 - IDENTIFICATION OF PHOSPHORYLATION SITES
PHOSPHORYLATION OF HUMAN SPERM PROTAMINES HP1 AND HP2 - IDENTIFICATION OF PHOSPHORYLATION SITES
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DOI：
10.1016/0167-4838(93)90043-q
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发表时间：
1993-11-10
期刊：
影响因子：
--
通讯作者：
SAUTIERE, P
中科院分区：
文献类型：
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作者：
CHIRAT, F;ARKHIS, A;SAUTIERE, P
人类精子的特征在于其鱼精蛋白原、鱼精蛋白和组蛋白的基本核蛋白补体的高度异质性。这种异质性增加了持久的磷酸化鱼精蛋白在成熟精子。碱性磷酸酶处理表明鱼精蛋白HP 1和HP 2被不同程度的磷酸化。通过电喷雾质谱法进一步证明了非磷酸化和磷酸化鱼精蛋白HP 1和HP 2的存在。在磷酸丝氨酸衍生为S-乙基半胱氨酸后，通过蛋白质的自动Edman降解来鉴定单磷酸化和二磷酸化鱼精蛋白HP 1的磷酸化位点。在两种磷酸化形式中，发现Ser-10被磷酸化;在二磷酸化形式中，Ser-8被鉴定为磷酸化的第二位点。在鱼精蛋白HP 2中，通过有限酸水解酶解肽和薄层电泳，定位了唯一的磷酸化位点（Ser-14）。
Human sperm is characterized by a high heterogeneity of its basic nuclear protein complement of pro-protamines, protamines and histones. This heterogeneity is increased by the persistence of phosphorylated protamines in mature spermatozoa. Alkaline phosphatase treatment of whole protein indicated that protamines HP1 and HP2 were phosphorylated to various degrees. Presence of non-phosphorylated and phosphorylated protamines HP1 and HP2 was further demonstrated by electrospray mass spectrometry. Phosphorylation sites of mono- and di-phosphorylated protamine HP1 were identified by automatic Edman degradation of the protein after phosphoserine derivatization to S-ethylcysteine. In both phosphorylated forms, Ser-10 was found phosphorylated; in the di-phosphorylated form, Ser-8 was identified as the second site of phosphorylation. In protamine HP2, the unique site of phosphorylation (Ser-14) was located after limited acid hydrolysis of enzymic peptides and thin-layer electrophoresis.