CRYSTALLIZATION AND PRELIMINARY-X-RAY STUDIES OF NADPH-CYTOCHROME P450 REDUCTASE

被引：30

作者：

DJORDJEVIC, S

ROBERTS, DL

WANG, M

SHEA, T

CAMITTA, MGW

MASTERS, BSS

KIM, JJP

机构：

[1] MED COLL WISCONSIN,DEPT BIOCHEM,MILWAUKEE,WI 53226

[2] UNIV TEXAS,HLTH SCI CTR,DEPT BIOCHEM,SAN ANTONIO,TX 78284

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1995年 / 92卷 / 08期

关键词：

X-RAY CRYSTALLOGRAPHY; FLAVOPROTEIN;

D O I：

10.1073/pnas.92.8.3214

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

NADPH-cytochrome P450 reductase (CPR; NADPH:ferrihemoprotein reductase, EC 1.6.2.4) catalyzes the transfer of electrons to all known microsomal cytochromes P450. CPR is unique in that it is one of only two mammalian enzymes known to contain both flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN), the other being the various isoforms of nitric oxide synthase. Similarities in amino acid sequence and in functional domain arrangement with other key flavoproteins, including nitric oxide synthase, make CPR an excellent prototype for studies of interactions between two flavin cofactors. We have obtained diffraction-quality crystals of rat liver CPR, expressed in Escherichia coli and solubilized by limited proteolysis with trypsin. The crystals were grown in Hepes buffer (pH 7.0), containing polyethylene glycol 4500 and NaCl. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit cell dimensions a = 103.3 Angstrom, b = 116.1 Angstrom, and c = 120.4 Angstrom. If we assume that there are two molecules of the 72-kDa CPR polypeptide per asymmetric unit, the calculated value of V-m is 2.54 Angstrom(3)/Da.

引用

页码：3214 / 3218

页数：5

共 49 条

[1]

Williams C.H., Kamin H., J. Biol. Chem., 237, pp. 587-595, (1962)

[2]

Phillips A.H., Langdon R.G., J. Biol. Chem., 237, pp. 2652-2660, (1962)

[3]

Schacter B.A., Nelson E.B., Marver H.S., Masters B.S.S., J. Biol. Chem., 247, pp. 3601-3607, (1972)

[4]

Horecker B.L., J. Biol. Chem., 183, pp. 593-605, (1950)

[5]

Enoch H.G., Strittmatter P., J. Biol. Chem., 254, pp. 8976-8981, (1979)

[6]

Vermilion J.L., Ballou D.P., Massey V., Coon M.J., J. Biol. Chem., 256, pp. 266-277, (1981)

[7]

Masters B.S.S., Bilimoria M.H., Kamin H., Gibson Q.H., J. Biol. Chem., 240, pp. 4081-4088, (1965)

[8]

Iyanagi T., Makino R., Anan F.K., Biochemistry, 20, pp. 1722-1730, (1981)

[9]

Guengerich F.P., Biochemistry, 22, pp. 2811-2820, (1983)

[10]

Otvos J.D., Krum D.P., Masters B.S.S., Biochemistry, 25, pp. 7220-7228, (1986)

← 1 2 3 4 5 →