Krell T, Coggins J R, Lapthorn A J
Department of Chemistry, University of Glasgow, Glasgow G12 8QQ, Scotland, UK.
J Mol Biol. 1998 May 22;278(5):983-97. doi: 10.1006/jmbi.1998.1755.
The three-dimensional structure of shikimate kinase from Erwinia chrysanthemi has been determined by multiple isomorphous replacement. Two models are presented: a high resolution 1.9 A model and a 2.6 A model which contains bound Mg-ADP. The enzyme is an alpha/beta protein consisting of a central sheet of five parallel beta-strands flanked by alpha-helices with overall topology similar to adenylate kinase. Evidence is presented that shikimate kinase undergoes major conformational changes on ligand binding. It resembles adenylate kinase in having a P-loop containing core structure and two flexible domains which undergo induced fit movement on substrate binding. The binding of Mg2+ in the active site of shikimate kinase involves direct interaction with two protein side-chains which is different from the situation found in adenylate kinase. Shikimate kinase has a readily identifiable Walker A-motif and a recognisable but modified Walker B-motif. Comparison of shikimate kinase to adenylate kinase has led to the identification of an adenine-binding motif (I/VDAXQ/NXP). Difference Fourier calculations have revealed the shikimate binding site which corresponds to the location of the AMP-binding site in adenylate kinase. A model for shikimate-binding is presented.
通过多同晶置换法确定了菊欧文氏菌莽草酸激酶的三维结构。给出了两个模型:一个高分辨率的1.9埃模型和一个包含结合态Mg-ADP的2.6埃模型。该酶是一种α/β蛋白,由一个由五条平行β链组成的中央片层以及两侧的α螺旋构成,其整体拓扑结构类似于腺苷酸激酶。有证据表明,莽草酸激酶在配体结合时会发生重大构象变化。它与腺苷酸激酶相似,具有一个含P环的核心结构和两个柔性结构域,在底物结合时会发生诱导契合运动。莽草酸激酶活性位点中Mg2+的结合涉及与两个蛋白质侧链的直接相互作用,这与腺苷酸激酶中的情况不同。莽草酸激酶有一个易于识别的沃克A基序和一个可识别但经过修饰的沃克B基序。将莽草酸激酶与腺苷酸激酶进行比较,从而确定了一个腺嘌呤结合基序(I/VDAXQ/NXP)。差值傅里叶计算揭示了莽草酸结合位点,该位点与腺苷酸激酶中AMP结合位点的位置相对应。给出了一个莽草酸结合模型。