Druck T, Gu Y, Prabhala G, Cannizzaro L A, Park S H, Huebner K, Keen J H
Jefferson Cancer Institute, Jefferson Medical College, Philadelphia, Pennsylvania 19107, USA.
Genomics. 1995 Nov 1;30(1):94-7. doi: 10.1006/geno.1995.0016.
Clathrin-coated vesicles, involved in endocytosis and Golgi processing, have a surface lattice containing clathrin triskelia and stoichiometric amounts of additional components termed "assembly proteins," or APs. The AP form at the plasma membrane, AP2, is composed of two large subunits of 100-115 kDa, denoted AP2 alpha and AP2 beta, a medium chain of 50 kDa, designated AP50, and a small chain. We have determined human chromosomal locations of genes for a large AP2 beta (CLAPB1) and a medium (CLAPM1) AP subunit and of a novel clathrin-binding protein, VCP, that binds clathrin simultaneously with APs. Chromosomal in situ hybridization of a human genomic clone demonstrated that the CLAPM1 gene mapped to chromosome region 3q28. The gene for the CLAPB1 large subunit was mapped to 17q11.2-q12 by PCR amplification of an AP2 beta fragment from a panel of rodent-human hybrid DNAs. To map the human VCP sequence, a human-specific probe was made by RT-PCR of human mRNA using oligonucleotide primers from conserved regions of the porcine sequence. The amplified human fragment served as probe on Southern blots of hybrid DNAs to determine that the human VCP locus maps to chromosome region 9pter-q34.
网格蛋白包被小泡参与内吞作用和高尔基体加工过程,其表面晶格含有网格蛋白三脚复合体以及化学计量的其他成分,这些成分被称为“装配蛋白”或AP。在质膜上形成的AP形式,即AP2,由两个100 - 115 kDa的大亚基(分别称为AP2α和AP2β)、一个50 kDa的中链(称为AP50)和一个小链组成。我们已经确定了一个大的AP2β(CLAPB1)和一个中等大小(CLAPM1)的AP亚基以及一种新型网格蛋白结合蛋白VCP(它能同时与AP结合网格蛋白)的基因在人类染色体上的位置。一个人类基因组克隆的染色体原位杂交表明,CLAPM1基因定位于染色体区域3q28。通过对一组啮齿动物 - 人类杂交DNA中的AP2β片段进行PCR扩增,将CLAPB1大亚基的基因定位于17q11.2 - q12。为了定位人类VCP序列,利用来自猪序列保守区域的寡核苷酸引物通过人类mRNA的RT - PCR制备了一个人类特异性探针。扩增得到的人类片段用作杂交DNA Southern印迹的探针,以确定人类VCP基因座定位于染色体区域9pter - q34。