Evidence for a Molten Globule State in Cicer α -Galactosidase Induced by pH, Temperature, and Guanidine Hydrochloride
作者： Neelesh Singh , Reetesh Kumar , M. V. Jagannadham , Arvind M. Kayastha
DOI: 10.1007/S12010-013-0163-9
关键词: Fluorescence spectroscopy 、 Denaturation (biochemistry) 、 Tryptophan 、 Chemistry 、 Crystallography 、 Guanidine 、 Molten globule 、 Enzyme 、 Circular dichroism 、 Protein secondary structure
摘要: Physiologically as well industrially, α-galactosidases are very important enzymes, but little is known about the stability and folding aspect of enzyme. In present study, we have investigated temperature, pH, guanidine hydrochloride (GuHCl) induced unfolding Cicer α-galactosidase using circular dichroism fluorescence spectroscopy. Strong negative ellipticities at 208, 215, 222 nm indicate presence both α β structures in showed that its secondary structure belongs to + class proteins with 31 % α-helicity. For emission maximum was found be 345 which suggests tryptophan residues less exposed solvent. However, pH 2.0, protein blue-shift. This state lacked activity it retained significant structure. Enhanced binding ANS 2.0 indicated exposure hydrophobic regions. The unfolded a red-shift 15 concomitant decrease intensity. enzyme maintained native full up 40 °C; however, above this denaturation observed.
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