Crystallization and preliminary crystallographic studies of the metalloglycoprotein esterase A4 using a baculovirus expression system

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 63; no. 9; pp. 734 - 736
• Main Authors: Hiraki, Toshiki, Shibayama, Naoya, Yoon, Young-Ho, Yun, Kyung-Mook, Hamamoto, Toshiro, Tame, Jeremy R. H., Park, Sam-Yong
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Blackwell Publishing Ltd 01-09-2007; International Union of Crystallography
• Subjects: Animals; Baculoviridae; Baculovirus; Base Sequence; Bombyx - enzymology; Bombyx mori; Carboxylesterase - chemistry; Carboxylesterase - isolation & purification; CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; Crystallization Communications; Crystallography, X-Ray; CRYSTALS; DIMERS; DNA Primers; esterase A4; Female; Glycoproteins - chemistry; Glycoproteins - isolation & purification; Glycosylation; Metalloproteins - chemistry; Metalloproteins - isolation & purification; Molecular Sequence Data; Ovum - enzymology; PROTEINS; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; RESOLUTION; SOLVENTS; SPACE GROUPS; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; SYNCHROTRON RADIATION; WEIGHT
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309107033854

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 Å resolution at 100 K using synchrotron radiation. The protein crystals belong to space group P21, with unit‐cell parameters a = 47.1, b = 73.9, c = 47.4 Å, β = 104.1°. With one dimer per asymmetric unit, the crystal volume per unit protein weight (VM) is 2.3 Å3 Da−1 and the solvent content is 47%.