Arribas J, Massagué J
Cell Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York 10021.
J Cell Biol. 1995 Feb;128(3):433-41. doi: 10.1083/jcb.128.3.433.
Cleavage and release of membrane protein ectodomains, a regulated process that affects many cell surface proteins, remains largely uncharacterized. To investigate whether cell surface proteins are cleaved through a shared mechanism or through multiple independent mechanisms, we mutagenized Chinese hamster ovary (CHO) cells and selected clones that were unable to cleave membrane-anchored transforming growth factor alpha (TGF-alpha). The defect in TGF-alpha cleavage in these clones is most apparent upon cell treatment with the protein kinase C (PKC) activator PMA, which stimulates TGF-alpha cleavage in wild-type cells. The mutant clones do not have defects in TFG-alpha expression, transport to the cell surface or turnover. Concomitant with the loss of TGF-alpha cleavage, these clones have lost the ability to cleave many structurally unrelated membrane proteins in response to PMA. These proteins include beta-amyloid precursor protein (beta-APP), whose cleavage into a secreted form avoids conversion into the amyloidogenic peptide A beta, and a group of cell surface proteins whose release into the medium is stimulated by PMA in wild type CHO cells but not in mutants. The mutations prevent cleavage by PKC-dependent as well as PKC-independent mechanisms, and thus affect an essential component that functions downstream of these various signaling mechanisms. We propose that regulated cleavage and secretion of membrane protein ectodomains is mediated by a common system whose components respond to multiple activators and act on susceptible proteins of diverse structure and function.
膜蛋白胞外域的切割与释放是一个影响许多细胞表面蛋白的调控过程,目前在很大程度上仍未得到充分研究。为了探究细胞表面蛋白是通过共享机制还是多种独立机制进行切割,我们对中国仓鼠卵巢(CHO)细胞进行了诱变,并筛选出无法切割膜锚定转化生长因子α(TGF-α)的克隆。在用蛋白激酶C(PKC)激活剂佛波酯(PMA)处理细胞时,这些克隆中TGF-α切割的缺陷最为明显,而PMA可刺激野生型细胞中的TGF-α切割。突变克隆在TFG-α表达、转运至细胞表面或周转方面没有缺陷。与TGF-α切割功能丧失相伴的是,这些克隆丧失了响应PMA切割许多结构不相关膜蛋白的能力。这些蛋白包括β-淀粉样前体蛋白(β-APP),其切割成分泌形式可避免转化为淀粉样生成肽Aβ,以及一组细胞表面蛋白,在野生型CHO细胞中,PMA可刺激它们释放到培养基中,但在突变体中则不然。这些突变阻止了通过PKC依赖性和PKC非依赖性机制的切割,因此影响了在这些各种信号传导机制下游起作用的一个关键组分。我们提出,膜蛋白胞外域的调控切割和分泌是由一个共同系统介导的,该系统的组分对多种激活剂作出反应,并作用于结构和功能各异的易感蛋白。