REFINED STRUCTURE FOR THE COMPLEX OF D-GLUCO-DIHYDROACARBOSE WITH GLUCOAMYLASE FROM ASPERGILLUS-AWAMORI VAR X100 TO 2.2-ANGSTROM RESOLUTION - DUAL CONFORMATIONS FOR EXTENDED INHIBITORS BOUND TO THE ACTIVE-SITE OF GLUCOAMYLASE

被引:68
作者
STOFFER, B
ALESHIN, AE
FIRSOV, LM
SVENSSON, B
HONZATKO, RB
机构
[1] CARLSBERG LAB,DEPT CHEM,DK-2500 COPENHAGEN,DENMARK
[2] ST PETERSBURG NUCL PHYS INST,DEPT MOLEC BIOL,GATCHINA 188350,RUSSIA
基金
美国国家科学基金会;
关键词
X-RAY CRYSTALLOGRAPHY; GLUCOAMYLASE STRUCTURE; ENZYME MECHANISM; OLIGOSACCHARIDE HYDROLYSIS; CARBOHYDRATE PROTEIN INTERACTION;
D O I
10.1016/0014-5793(94)01354-4
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The crystal structure at pH 4 of the complex of glucoamylase II(471) from Aspergillus awamori var. X100 with the pseudotetrasaccharide D-gluco-dihydroacarbose has been refined to an R-factor of 0.125 against data to 2.2 Angstrom resolution. The first two residues of the inhibitor bind at a position nearly identical to those of the closely related inhibitor acarbose in its complex with glucoamylase at pH 6. However, the electron density bifurcates beyond the second residue of the D-gluco-dihydroacarbose molecule, placing the third and fourth residues together at two positions in the active site. The position of relatively low density (estimated occupancy of 35%) corresponds to the location of the third and fourth residues of acarbose in its complex with glucoamylase at pH 6. The position of high density (65% occupancy) corresponds to a new binding mode of an extended inhibitor to the active site of glucoamylase. Presented are possible causes for the binding of D-gluco-dihydroacarbose in two conformations at the active site of glucoamylase at pH 4.
引用
收藏
页码:57 / 61
页数:5
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