Analysis of the Heat-Induced Denaturation of Proteins Using Temperature Gradient Gel Electrophoresis

✍ Scribed by T. Arakawa; L. Hung; V. Pan; T.P. Horan; C.G. Kolvenbach; L.O. Narhi

• Publisher: Elsevier Science
• Year: 1993
• Tongue: English
• Weight: 486 KB
• Volume: 208
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1993.1042

✦ Synopsis

A new technique, temperature gradient gel electrophoresis (TGGE), was applied to the study of heat-induced protein denaturation. The gels used contained 30 mм Borax (+75 \mathrm{~mm}) boric acid, (\mathrm{pH}) 8.4, and various concentrations of urea. When this technique was applied to bovine serum albumin, the protein showed discontinuous bands upon melting, indicating that the thermal transition is irreversible. The apparent melting temperature, calculated based on the relative intensity of two bands in the transition region, was (58^{\circ} \mathrm{C}) in (2 \mathrm{~m}) urea. When the thermal denaturation of bovine serum albumin was analyzed spectroscopically, the transition was again irreversible, with an apparent transition temperature of (57^{\circ} \mathrm{C}), consistent with the TGGE results. Recombinant stem cell factor, recombinant granulocyte macrophage-colony stimulating factor, and catalase were also analyzed by TGGE, indicating that the technique can be used to analyze denaturation of monomeric and multimeric proteins. 1983 Academic Press, Inc.