کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5512346 | 1540224 | 2017 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Purification, preliminary X-ray crystallography and biophysical studies of triose phosphate isomerase-β-globin subunit complex
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Purification, preliminary X-ray crystallography and biophysical studies of triose phosphate isomerase-β-globin subunit complex Purification, preliminary X-ray crystallography and biophysical studies of triose phosphate isomerase-β-globin subunit complex](/preview/png/5512346.png)
چکیده انگلیسی
Triose phosphate isomerase (TIM) is a cytoplasmic enzyme of prime importance in the mammalian glycolytic pathway. It has a major role in the conversion of dihydroxyacetone phosphate into glyceraldehyde-3-phosphate. We have successfully purified a stable complex of TIM with β-globin subunit from the sheep kidney using a simple two-step chromatography procedure. It is seen for the first time that TIM is forming a stable complex with β-globin. The purified protein-protein complex was crystallized and preliminary diffraction data were collected at 2.1 Ã
resolution. We further studied guanidinium chloride (GdmCl)-induced denaturation of TIM-β-globin complex by monitoring changes in the mean residue ellipticity at 222 nm ([θ]222) and difference absorption coefficient at 406 nm (Îε406) at pH 7.5 and 25 °C. We have observed that GdmCl-induced denaturation is reversible. Coincidence of normalized transition curves of both physical properties ([θ]222 and Îε406) suggests that folding/unfolding of TIM and β-subunit proteins is a two-state process. Denaturation curves of [θ]222 and Îε406 were used to estimate the stability parameters of the protein-protein complex. This is the first report on the isolation, purification, crystallization and biophysical characterization of the naturally occurring complex of TIM with the β-globin subunit.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Biological Macromolecules - Volume 94, Part B, January 2017, Pages 746-753
Journal: International Journal of Biological Macromolecules - Volume 94, Part B, January 2017, Pages 746-753
نویسندگان
Wahiduzzaman Wahiduzzaman, Mohammad Aasif Dar, Mohd. Amir, Asimul Islam, Md. Imtaiyaz Hassan, Faizan Ahmad,