Vogel B E, Lee S J, Hildebrand A, Craig W, Pierschbacher M D, Wong-Staal F, Ruoslahti E
La Jolla Cancer Research Foundation, California 92037.
J Cell Biol. 1993 Apr;121(2):461-8. doi: 10.1083/jcb.121.2.461.
Several studies have addressed the interaction of the HIV Tat protein with the cell surface. Our analysis of the cell attachment-promoting activity of Tat and peptides derived from it revealed that the basic domain of Tat, not the arg-gly-asp (RGD) sequence, is required for cell attachment to Tat. Affinity chromatography with Tat peptides and immunoprecipitation with various anti-integrin antibodies suggest that the vitronectin-binding integrin, alpha v beta 5, is the cell surface protein that binds to the basic domain of Tat. The Tat basic domain contains the sequence RKKRRQRRR. A related sequence, KKQRFRHRNRKG, present in the heparin-binding domain of an alpha v beta 5 ligand, vitronectin, also bound alpha v beta 5 in affinity chromatography and, in combination with an RGD peptide, was an inhibitor of cell attachment to vitronectin. The alpha v beta 5 interaction with these peptides was not solely due to high content of basic amino acids in the ligand sequences; alpha v beta 5 did not bind substantially to peptides consisting entirely of arginine or lysine, whereas a beta 1 integrin did bind to these peptides. The interaction of alpha v beta 5 with Tat is atypical for integrins in that the binding to Tat is divalent cation independent, whereas the binding of the same integrin to an RGD-containing peptide or to vitronectin requires divalent cations. These data define an auxiliary integrin binding specificity for basic amino acid sequences. These basic domain binding sites may function synergistically with the binding sites that recognize RGD or equivalent sequences.
多项研究探讨了HIV反式激活因子(Tat蛋白)与细胞表面的相互作用。我们对Tat及其衍生肽的细胞附着促进活性的分析表明,Tat的碱性结构域而非精氨酸-甘氨酸-天冬氨酸(RGD)序列是细胞附着于Tat所必需的。用Tat肽进行亲和层析以及用各种抗整合素抗体进行免疫沉淀表明,玻连蛋白结合整合素αvβ5是与Tat碱性结构域结合的细胞表面蛋白。Tat碱性结构域包含序列RKKRRQRRR。一个相关序列KKQRFRHRNRKG存在于αvβ5配体玻连蛋白的肝素结合结构域中,在亲和层析中也能结合αvβ5,并且与RGD肽结合时,是细胞附着于玻连蛋白的抑制剂。αvβ5与这些肽的相互作用并非仅仅归因于配体序列中碱性氨基酸的高含量;αvβ5与完全由精氨酸或赖氨酸组成的肽基本不结合,而β1整合素则能与这些肽结合。αvβ5与Tat的相互作用对于整合素来说是非典型的,因为其与Tat的结合不依赖二价阳离子,而同一整合素与含RGD肽或玻连蛋白的结合则需要二价阳离子。这些数据定义了整合素对碱性氨基酸序列的一种辅助结合特异性。这些碱性结构域结合位点可能与识别RGD或等效序列的结合位点协同发挥作用。