Crystallization and preliminary crystallographic studies of the metalloglycoprotein esterase A4 using a baculovirus expression system.
Esterase A4 (EA4) is a timer protein found in diapause eggs of the silkworm Bombyx mori. The gene for this metalloglycoprotein was cloned from B. mori eggs and expressed using a baculovirus expression system in silkworm pupae. Crystals of the purified protein have been grown that diffract to beyond 2.1 A resolution at 100 K using synchrotron radiation. The protein crystals belong to space group P2(1), with unit-cell parameters a = 47.1, b = 73.9, c = 47.4 A, beta = 104.1 degrees. With one dimer per asymmetric unit, the crystal volume per unit protein weight (V(M)) is 2.3 A3 Da(-1) and the solvent content is 47%.
Hiraki
Toshiki
T
Protein Design Laboratory, Yokohama City University, 1-7-29 Suehiro, Tsurumi, Yokohama 230-0045, Japan.
Shibayama
Naoya
N
Yoon
Young-Ho
YH
Yun
Kyung-Mook
KM
Hamamoto
Toshiro
T
Tame
Jeremy R H
JR
Park
Sam-Yong
SY
D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D012831:Bombyx / Q000201:enzymology; D043182:Carboxylesterase / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D005260:Female; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification; D006031:Glycosylation; D008667:Metalloproteins / Q000737:chemistry / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010063:Ovum / Q000201:enzymology; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D017931:DNA Primers; D006023:Glycoproteins; D008667:Metalloproteins; D011994:Recombinant Proteins; D043182:Carboxylesterase