Fry D C, Kuby S A, Mildvan A S
Proc Natl Acad Sci U S A. 1986 Feb;83(4):907-11. doi: 10.1073/pnas.83.4.907.
The MgATP binding site of adenylate kinase, located by a combination of NMR and x-ray diffraction, is near three protein segments, five to seven amino acids in length, that are homologous in sequence to segments found in other nucleotide-binding phosphotransferases, such as myosin and F1-ATPase, ras p21 and transducin GTPases, and cAMP-dependent and src protein kinases, suggesting equivalent mechanistic roles of these segments in all of these proteins. Segment 1 is a glycine-rich flexible loop that, on adenylate kinase, may control access to the ATP-binding site by changing its conformation. Segment 2 is an alpha-helix containing two hydrophobic residues that interact with the adenine-ribose moiety of ATP, and a lysine that may bind to the beta- and gamma-phosphates of ATP. Segment 3 is a hydrophobic strand of parallel beta-pleated sheet, terminated by a carboxylate, that flanks the triphosphate binding site. The various reported mutations of ras p21 that convert it to a transforming agent all appear to involve segment 1, and such substitutions may alter the properties of p21 by hindering a conformational change at this segment. In F1-ATPase, the flexible loop may, by its position, control both the accessibility and the ATP/ADP equilibrium constant on the enzyme.
通过核磁共振(NMR)和X射线衍射相结合的方法确定,腺苷酸激酶的MgATP结合位点靠近三个蛋白质片段,每个片段长度为五到七个氨基酸,它们在序列上与其他核苷酸结合磷酸转移酶中的片段同源,如肌球蛋白和F1 - ATP酶、ras p21和转导素GTP酶,以及cAMP依赖性蛋白激酶和src蛋白激酶,这表明这些片段在所有这些蛋白质中具有相同的机制作用。片段1是一个富含甘氨酸的柔性环,在腺苷酸激酶上,它可能通过改变构象来控制对ATP结合位点的 access。片段2是一个α螺旋,包含两个与ATP的腺嘌呤 - 核糖部分相互作用的疏水残基,以及一个可能与ATP的β和γ磷酸基团结合的赖氨酸。片段3是一条由羧酸盐终止的平行β折叠片层的疏水链,位于三磷酸结合位点的两侧。所有已报道的将ras p21转化为转化剂的突变似乎都涉及片段1,这种取代可能通过阻碍该片段的构象变化来改变p21的性质。在F1 - ATP酶中,柔性环可能通过其位置控制酶上的 accessibility和ATP/ADP平衡常数。 (注:原文中“access”翻译时结合语境推测为“接近、进入”之类意思,但不确定准确含义,这里保留原文以便进一步确认)