Bennett W S, Steitz T A
Proc Natl Acad Sci U S A. 1978 Oct;75(10):4848-52. doi: 10.1073/pnas.75.10.4848.
The A isozyme of yeast hexokinase (ATP:D-hexose 6-phosphotransferase, EC 2.7.1.1) crystallized as a complex with glucose has a conformation that is dramatically different from the conformation of the B isozyme crystallized in the absence of glucose. Comparison of the high-resolution structures shows that one lobe of the molecule is rotated by 12 degrees relative to the other lobe, resulting in movements of as much as 8 A in the polypeptide backbone and closing the cleft between the lobes into which glucose is bound. The conformational change is produced by the binding of glucose (R.C. McDonald, T.A. Steitz, and D.M. Engelman, unpublished data) and is essential for catalysis [Anderson, C.M., Stenkamp, R.E., McDonald, R.C. & Steitz, T.A. (1978) J. Mol. Biol. 123, 207-219] and thus provides an example of induced fit. The surface area of the hexokinase A-glucose complex exposed to solvent is smaller than that of native hexokinase B. By using the change in exposed surface area to estimate the hydrophobic contribution to the free energy changes upon glucose binding, we find that the hydrophobic effect alone favors the active conformation of hexokinase in the presence and absence of sugar. The observed stability of the inactive conformation of the enzyme in the absence of substrates may result from a deficiency of complementary interactions within the cavity that forms when the two lobes close together.
酵母己糖激酶的A同工酶(ATP:D-己糖6-磷酸转移酶,EC 2.7.1.1)与葡萄糖形成复合物结晶时的构象,与在无葡萄糖情况下结晶的B同工酶的构象显著不同。高分辨率结构的比较表明,分子的一个叶相对于另一个叶旋转了12度,导致多肽主链移动多达8埃,并封闭了结合葡萄糖的叶之间的裂隙。这种构象变化是由葡萄糖的结合产生的(R.C.麦克唐纳、T.A.施泰茨和D.M.恩格尔曼,未发表数据),并且对于催化作用至关重要[安德森,C.M.,斯滕坎普,R.E.,麦克唐纳,R.C.和施泰茨,T.A.(1978年)《分子生物学杂志》123卷,207 - 219页],因此提供了一个诱导契合的例子。己糖激酶A - 葡萄糖复合物暴露于溶剂的表面积比天然己糖激酶B的小。通过利用暴露表面积的变化来估计葡萄糖结合时对自由能变化的疏水贡献,我们发现仅疏水效应在有糖和无糖情况下都有利于己糖激酶的活性构象。在没有底物时观察到的酶的无活性构象的稳定性,可能是由于当两个叶靠近时形成的腔内互补相互作用不足所致。