Expression, purification, crystallization and preliminary X-ray analysis of perakine reductase, a new member of the aldo-keto reductase enzyme superfamily from higher plants.
Perakine reductase (PR) is a novel member of the aldo-keto reductase enzyme superfamily from higher plants. PR from the plant Rauvolfia serpentina is involved in the biosynthesis of monoterpenoid indole alkaloids by performing NADPH-dependent reduction of perakine, yielding raucaffrinoline. However, PR can also reduce cinnamic aldehyde and some of its derivatives. After heterologous expression of a triple mutant of PR in Escherichia coli, crystals of the purified and methylated enzyme were obtained by the hanging-drop vapour-diffusion technique at 293 K with 100 mM sodium citrate pH 5.6 and 27% PEG 4000 as precipitant. Crystals belong to space group C222(1) and diffract to 2.0 A, with unit-cell parameters a = 58.9, b = 93.0, c = 143.4 A.
Rosenthal
Cindy
C
Department of Pharmaceutical Biology, Institute of Pharmacy, Johannes Gutenberg-University Mainz, Staudinger Weg 5, D-55099 Mainz, Germany.
Mueller
Uwe
U
Panjikar
Santosh
S
Sun
Lianli
L
Ruppert
Martin
M
Zhao
Yu
Y
Stöckigt
Joachim
J
D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D003460:Crystallization; D018360:Crystallography, X-Ray; D011926:Rauwolfia / Q000201:enzymology*
D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase