Heat-shock cognate protein (hsc71) and related proteins in mouse spermatogenic cells.
Heat-shock cognate protein (hsc71) and related proteins in mouse spermatogenic cells.
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小鼠生精细胞中的热休克同源蛋白 (hsc71) 和相关蛋白。
DOI：
10.1095/biolreprod40.4.843
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发表时间：
1989
影响因子：
3.6
通讯作者：
E. M. Eddy
中科院分区：
文献类型：
--
作者：
M. Maekawa;D. A. O’Brien;R. Allen;E. M. Eddy
已经开发了一种单克隆抗体（13 D3），其识别从小鼠生精细胞（主要是粗线期精母细胞和精子细胞）的混合物中提取的蛋白质的二维免疫印迹上的71千道尔顿（71 kDa）蛋白质。该蛋白质通过免疫印迹和三磷酸腺苷（ATP）结合特性显示与存在于3 T3细胞中的71 kDa小鼠热休克同源（HSC）蛋白hsc 71相同。沿着70 kDa热休克诱导蛋白（hsp 70）和74 kDa热休克同源蛋白（hsc 74），hsc 71是小鼠HSP 70多基因家族的产物。尽管抗体13 D3与hsc 71强烈反应，但在3 T3细胞中它仅与hsp 70微弱反应，并且在混合生殖细胞的免疫印迹上完全不与hsc 74或生殖细胞特异性hsp 70样蛋白（P70）反应。抗体13 D3在已知抗体中与这些热休克蛋白的反应模式是独特的。在分离的生殖细胞的免疫荧光研究中，13 D3反应均匀的粗线期精母细胞，圆形精子细胞，和残留的机构的细胞质，但只与精子的中段。抗体13 D3识别其他蛋白质，除了hsc 71的二维免疫印迹凝聚精子细胞和精子。其中两种蛋白质（70 kDa/pI6.4和70 kDa/pI6.5）存在于凝聚精子细胞和精子中，另一种蛋白质（69 kDa/pI7.0）仅在精子中检测到。新蛋白也被单克隆抗体7.10识别，其与hsp 70、hsc 71、hsc 74和P70特异性反应。虽然[35 S]蛋氨酸被纳入新的蛋白质在凝聚精子细胞，hsc 71，hsc 74，和P70没有标记。这些结果表明，独特的热休克蛋白在精子发生后期合成。
A monoclonal antibody (13D3) has been developed that recognizes a 71 kilodalton (71 kDa) protein on two-dimensional immunoblots of proteins extracted from a mixture of mouse spermatogenic cells (mainly pachytene spermatocytes and spermatids). This protein was shown by immunoblotting and adenosine triphosphate (ATP)-binding characteristics to be identical to a 71 kDa mouse heat-shock cognate (hsc) protein, hsc71, present in 3T3 cells. Along with a 70 kDa heat-shock inducible protein (hsp70), and a 74 kDa heat-shock cognate protein (hsc74), hsc71 is a product of the mouse HSP70 multigene family. Although antibody 13D3 reacted strongly with hsc71, it reacted only faintly with hsp70 in 3T3 cells, and not at all with hsc74 or a germ cell-specific hsp70-like protein (P70) on immunoblots of mixed germ cells. Antibody 13D3 is unique among known antibodies in its pattern of reaction with these heat-shock proteins. In immunofluorescence studies on isolated germ cells, 13D3 reacted uniformly with the cytoplasm of pachytene spermatocytes, round spermatids, and residual bodies, but only with the midpiece of spermatozoa. Antibody 13D3 recognizes other proteins in addition to hsc71 on two-dimensional immunoblots of condensing spermatids and spermatozoa. Two of the proteins (70 kDa/pI 6.4 and 70 kDa/pI 6.5) were present in condensing spermatids and spermatozoa, and another protein (69 kDa/pI 7.0) was detected only in spermatozoa. The new proteins also were recognized by monoclonal antibody 7.10, which reacts specifically with hsp70, hsc71, hsc74, and P70. Although [35S]methionine was incorporated into the new proteins in condensing spermatids, hsc71, hsc74, and P70 were not labeled. These results suggest that unique heat-shock proteins are synthesized late in spermatogenesis.
哺乳动物 70,000 道尔顿应激蛋白的快速纯化：蛋白质对核苷酸的亲和力。
影响因子：
5.3
作者：
Welch,WJ;Feramisco,JR
通讯作者：
Feramisco,JR
影响因子：
56.9
作者：
KURTZ, S;ROSSI, J;LINDQUIST, S
通讯作者：
LINDQUIST, S