SYNTHESIS AND AMINO-ACID COMPOSITION OF BASIC-PROTEINS IN MAMMALIAN SPERM NUCLEI
SYNTHESIS AND AMINO-ACID COMPOSITION OF BASIC-PROTEINS IN MAMMALIAN SPERM NUCLEI
复制标题
DOI：
10.1016/0012-1606(75)90289-4
复制
发表时间：
1975-01-01
影响因子：
2.7
通讯作者：
HANLEYBOWDOIN, L
中科院分区：
文献类型：
--
作者：
BELLVE, AR;ANDERSON, E;HANLEYBOWDOIN, L
采用聚丙烯酰胺凝胶电泳和氨基酸分析对人、鼠、兔和豚鼠精子核的基本染色体蛋白（SCP）进行了表征。用1% SDS去头精子，密度梯度离心恢复细胞核。诺玛斯基和电子显微镜检查显示细胞核完整，纯度达99%。从细胞核中提取碱性蛋白，经氨基乙基化，并用离子交换层析和凝胶过滤层析纯化。人、兔和豚鼠的SCP经聚丙烯酰胺凝胶电泳后，呈现出具有相似迁移率的单蛋白条带。相比之下，氨基化小鼠SCP由SCP·ae1和SCP·AE2两种蛋白组成，它们具有不同的电泳迁移率。这些哺乳动物的SCP富含精氨酸（47 ~ 54.4%）和半胱氨酸（7.7 ~ 12.2%）。这些蛋白质的氨基酸组成存在主要差异。小鼠和人SCP富含组氨酸（分别为12.2%和7.7%），豚鼠SCP富含酪氨酸（11.7%）和苯丙氨酸（3.5%）。仅在兔SCP中检测到缬氨酸，在人和豚鼠中检测到脯氨酸。4种植物均未检出天冬氨酸、蛋氨酸和色氨酸。对[3H]精氨酸掺入小鼠SCP的研究表明，这些基本蛋白是在精子发生的末期合成的，随后被保存下来。
The basic chromosomal proteins (SCP) of human, mouse, rabbit and guinea pig sperm nuclei were characterized by polyacrylamide gel electrophoresis and amino acid analysis. Spermatozoa were decapitated with 1% SDS and the nuclei recovered by density gradient centrifugation. Examination by Nomarski and electron microscopy revealed the nuclei to be intact and 99% pure. The basic proteins were extracted from nuclei, aminoethylated and purified by ion exchange chromatography and gel filtration chromatography.The SCP of human, rabbit and guinea pig gave single protein bands with similar mobilities when subjected to polyacrylamide gel electrophoresis. In contrast, aminoethylated mouse SCP consisted of two proteins, SCP·AE1and SCP·AE2, which had different electrophoretic mobilities. The SCP of these mammalian species were characteristically rich in arginine (47–54.4%) and cysteine (7.7–12.2%). Major differences existed in the amino acid compositions of these proteins. Mouse and human SCP were rich in histidine (12.2 and 7.7%, respectively) and guinea pig was high in tyrosine (11.7%) and phenylalanine (3.5%). Valine was detected only in rabbit SCP and proline in human and guinea pig. Aspartic acid, methionine and tryptophan were not detected in all four species. Studies on the incorporation of [3H]arginine into mouse SCP demonstrated that these basic proteins are synthesized during the terminal stages of spermatogenesis and are subsequently conserved.