Crystallization and preliminary X-ray analysis of crustacean hyperglycaemic hormone from the kuruma prawn Marsupenaeus japonicus in its weakly active precursor form

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Dec 1;67(Pt 12):1586-9. doi: 10.1107/S1744309111040140. Epub 2011 Nov 26.

Abstract

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 P2(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%.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arthropod Proteins / chemistry*
  • Crystallization
  • Crystallography, X-Ray
  • Invertebrate Hormones / chemistry*
  • Nerve Tissue Proteins / chemistry*
  • Penaeidae / chemistry*

Substances

  • Arthropod Proteins
  • Invertebrate Hormones
  • Nerve Tissue Proteins
  • hyperglycemic hormone, crustacean