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image of Purification, Crystallization, and Preliminary X-ray Diffraction Studies on Hemoglobin from the Angora Goat (Capra Aegagrus Hircus)

Abstract

Introduction

Angora goats are a distinct breed that differs significantly from common goats and shares a similar appearance to sheep. In Angora goats, only the level of glutathione (GSH) is elevated during under-stimulated conditions, as well as after the period of hypoxic stress; however, no changes are found in 2,3-diphosphoglycerate (2,3-DPG) levels, which are commonly present in the red blood cells (RBCs) of most mammals. We chose the Angora goat for our investigation because no previous studies have been conducted on the structural and functional aspects of hemoglobin (Hb). In addition, no sequence or structural information is currently available in any database.

Methods

Angora goat Hb was isolated and purified by anion-exchange chromatography, followed by crystallization using various methods. X-ray data collection for Angora goat Hb was performed under a liquid nitrogen cryo-stream using a system.

Results

Good diffracting crystals were obtained using the hanging-drop vapor-diffusion method with polyethylene glycol (PEG) 3350 as the precipitant in water, without the addition of any salt or buffer. The Angora goat Hb diffracted to a resolution of 1.85 Å, and the structure solution was obtained by the molecular replacement method, using the structure of domestic goat Hb as the starting model.

Discussion

The solved structure of Angora goat crystallized in the monoclinic space group P2, consisting of one whole biological molecule in the asymmetric unit, with unit cell dimensions of a = 52.08 Å, b = 76.70 Å, c = 74.08 Å, and β = 91.77 °. The solvent content and Matthews coefficient (Vm) for the Angora goat Hb are 49.05% and 2.41 Å3/Da, respectively, and are within the normal range for protein crystals.

Conclusion

Purification, crystallization, and preliminary X-ray diffraction studies of Angora goat Hb were performed successfully. Structural refinement and biophysical characterization of Angora goat Hb are in progress in the absence and presence of GSH and 2,3-DPG.

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2025-09-13

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Purification, Crystallization, and Preliminary X-ray Diffraction Studies on Hemoglobin from the Angora Goat (Capra Aegagrus Hircus)
Bentham Science Publishers ; https://doi.org/10.2174/0109298665370730250708065019
/content/journals/ppl/10.2174/0109298665370730250708065019
/content/journals/ppl/10.2174/0109298665370730250708065019
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  • Article Type:     Research Article
Keywords: hypoxic stress ; protein crystals ; oxygen affinity ; Angora goat ; level of glutathione ; hemoglobin

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