Open AccessCrystallization and preliminary X‐ray analysis of crustacean hyperglycaemic hormone from the kuruma prawn Marsupenaeus japonicus in its weakly active precursor form
Author(s) -
Inoue Hirotaka,
Tsutsui Naoaki,
Nagai Chiaki,
Nagata Koji,
Tanokura Masaru,
Nagasawa Hiromichi
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111040140
Subject(s) - prawn , orthorhombic crystal system , crystallization , biology , crustacean , escherichia coli , biochemistry , glycine , recombinant dna , chemistry , chromatography , crystallography , crystal structure , amino acid , fishery , gene , organic chemistry
Crustacean hyperglycaemic hormone (CHH) plays a pivotal role in the regulation of glucose metabolism in crustaceans. Pej‐SGP‐I, one of the six known CHHs in the kuruma prawn Marsupenaeus japonicus , was heterologously expressed in Escherichia coli as an N‐terminally His‐tagged and Nus‐tagged protein in its weakly active precursor form, Pej‐SGP‐I‐Gly, which has an extra glycine residue at the C‐terminus. The recombinant peptide was subjected to affinity purification, tag removal, further purification and crystallization by the sitting‐drop vapour‐diffusion method using NaCl as the main precipitant. The crystals diffracted to 1.95 Å resolution and the space group was assigned as primitive orthorhombic P 2 1 2 1 2 1 , with unit‐cell parameters a = 40.19, b = 53.65, c = 53.63 Å. The Matthews coefficient ( V M = 1.73 Å 3 Da −1 ) indicated that the crystal contained two Pej‐SGP‐I‐Gly molecules per asymmetric unit, with a solvent content of 29.0%.