Purification, crystallization and preliminary X-ray diffraction studies on goat (Capra hircus) hemoglobin - A low oxygen affinity species

Ponnuraj Sathya Moorthy, Kamariah Neelagandan, Balasubramanian Moovarkumudalvan, Mondikalipudur Nanjappa Gounder Ponnuswamy

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Hemoglobin is a vital protein present in almost all higher species. It is a transport protein involved in carrying oxygen from lungs to tissues and carbon dioxide back to lungs by an intrinsically coordinated manner. Even though a good amount of work has been carried out in this direction there exists scarcity of structural insight on low oxygen affinity species. Attempts are being made to unravel the structural insight of this low oxygen affinity species. Goat blood plasma was collected, treated with EDTA to avoid blood clotting and purification was accomplished using DEAE-anion chromatographic column. The goat hemoglobin was crystallized using 50mM of phosphate buffer at pH 6.7 with 1M NaCl and PEG 3350 as precipitant by hanging drop vapor diffusion method. Crystals obtained are screened and suitable crystals are taken for data collection using mar345dtb as image plate detector system. Goat hemoglobin crystal diffracted up to 2.61 Å resolution. Goat hemoglobin crystallizes in orthorhombic space group P212121 as a whole biological molecule in the asymmetric unit with cell dimensions a=53.568Å, b=67.365Å, c=154.183Å.

Original languageEnglish
Pages (from-to)454-456
Number of pages3
JournalProtein and Peptide Letters
Volume16
Issue number4
DOIs
Publication statusPublished - 1 Apr 2009
Externally publishedYes

Fingerprint

Crystallization
Goats
X-Ray Diffraction
Purification
Hemoglobins
Oxygen
X ray diffraction
Crystals
Blood
Lung
Distillation columns
Blood Coagulation
Carbon Dioxide
Edetic Acid
Anions
Carrier Proteins
Buffers
Phosphates
Vapors
Tissue

Keywords

  • 2, 3 disphosphoglycerate
  • Allosteric mechanism
  • Capra hircus
  • Chromatography
  • Low oxygen affinity
  • Orthorhombic

ASJC Scopus subject areas

  • Biochemistry
  • Structural Biology

Cite this

Purification, crystallization and preliminary X-ray diffraction studies on goat (Capra hircus) hemoglobin - A low oxygen affinity species. / Moorthy, Ponnuraj Sathya; Neelagandan, Kamariah; Moovarkumudalvan, Balasubramanian; Ponnuswamy, Mondikalipudur Nanjappa Gounder.

In: Protein and Peptide Letters, Vol. 16, No. 4, 01.04.2009, p. 454-456.

Research output: Contribution to journalArticle

Moorthy, Ponnuraj Sathya ; Neelagandan, Kamariah ; Moovarkumudalvan, Balasubramanian ; Ponnuswamy, Mondikalipudur Nanjappa Gounder. / Purification, crystallization and preliminary X-ray diffraction studies on goat (Capra hircus) hemoglobin - A low oxygen affinity species. In: Protein and Peptide Letters. 2009 ; Vol. 16, No. 4. pp. 454-456.
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