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

Hirotaka Inoue, Naoaki Tsutsui, Chiaki Nagai, Koji Nagata, Masaru Tanokura, Hiromichi Nagasawa

Research output: Contribution to journalArticle

5 Citations (Scopus)

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 heterolo-gously 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 12 12 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%.

Original languageEnglish
Pages (from-to)1586-1589
Number of pages4
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume67
Issue number12
DOIs
Publication statusPublished - Dec 2011
Externally publishedYes

Fingerprint

hormones
X ray analysis
Crystallization
purification
Glycine
Purification
X-Rays
crystallization
Escherichia coli
Glucose
Peptides
Crystals
metabolism
glycine
Escherichia
Metabolism
glucose
crystals
peptides
affinity

Keywords

  • crustacean hyperglycaemic hormone
  • Marsupenaeus japonicus

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Structural Biology
  • Genetics
  • Condensed Matter Physics

Cite this

Crystallization and preliminary X-ray analysis of crustacean hyperglycaemic hormone from the kuruma prawn Marsupenaeus japonicus in its weakly active precursor form. / Inoue, Hirotaka; Tsutsui, Naoaki; Nagai, Chiaki; Nagata, Koji; Tanokura, Masaru; Nagasawa, Hiromichi.

In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 67, No. 12, 12.2011, p. 1586-1589.

Research output: Contribution to journalArticle

@article{d7e1860b00f248feab75943ea856c5fe,
title = "Crystallization and preliminary X-ray analysis of crustacean hyperglycaemic hormone from the kuruma prawn Marsupenaeus japonicus in its weakly active precursor form",
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 heterolo-gously 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 {\AA} resolution and the space group was assigned as primitive orthorhombic P2 12 12 1, with unit-cell parameters a = 40.19, b = 53.65, c = 53.63 {\AA}. The Matthews coefficient (V M = 1.73 {\AA} 3 Da -1) indicated that the crystal contained two Pej-SGP-I-Gly molecules per asymmetric unit, with a solvent content of 29.0{\%}.",
keywords = "crustacean hyperglycaemic hormone, Marsupenaeus japonicus",
author = "Hirotaka Inoue and Naoaki Tsutsui and Chiaki Nagai and Koji Nagata and Masaru Tanokura and Hiromichi Nagasawa",
year = "2011",
month = "12",
doi = "10.1107/S1744309111040140",
language = "English",
volume = "67",
pages = "1586--1589",
journal = "Acta Crystallographica Section F:Structural Biology Communications",
issn = "1744-3091",
publisher = "John Wiley and Sons Ltd",
number = "12",

}

TY - JOUR

T1 - Crystallization and preliminary X-ray analysis of crustacean hyperglycaemic hormone from the kuruma prawn Marsupenaeus japonicus in its weakly active precursor form

AU - Inoue, Hirotaka

AU - Tsutsui, Naoaki

AU - Nagai, Chiaki

AU - Nagata, Koji

AU - Tanokura, Masaru

AU - Nagasawa, Hiromichi

PY - 2011/12

Y1 - 2011/12

N2 - 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 heterolo-gously 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 12 12 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%.

AB - 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 heterolo-gously 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 12 12 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%.

KW - crustacean hyperglycaemic hormone

KW - Marsupenaeus japonicus

UR - http://www.scopus.com/inward/record.url?scp=83055169803&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=83055169803&partnerID=8YFLogxK

U2 - 10.1107/S1744309111040140

DO - 10.1107/S1744309111040140

M3 - Article

C2 - 22139173

AN - SCOPUS:83055169803

VL - 67

SP - 1586

EP - 1589

JO - Acta Crystallographica Section F:Structural Biology Communications

JF - Acta Crystallographica Section F:Structural Biology Communications

SN - 1744-3091

IS - 12

ER -