title: Molten globule-like folding intermediate of asialofetuin at acidic pH.
abstract: In our earlier communication on acid-induced unfolding of bovine serum fetuin (BSF), we showed the existence of a molten globule (MG)-like state of BSF at pH 1.8. The MG state was characterized by higher content of secondary structure than native and almost complete loss of tertiary structure and more solvent exposed hydrophobic surface [Biochim. Biophys. Acta 1649 (2003) 164]. In this work we have shown the presence of an MG-like partially folded intermediate of asialofetuin at around pH 1.8, which is much different from the MG state observed in BSF in secondary structure contents. The results show that asialofetuin at pH 1.8 retains approximately 45% secondary structure, as evident from far-UV CD spectra. The near-UV CD spectra showed almost complete loss of tertiary structure. The intrinsic fluorescence and acrylamide quenching of the lone tryptophan residue showed that in acid-induced state, it is buried in the interior in a nonpolar environment. The temperature dependence of far-UV CD signal of asialofetuin at pH 1.8 exhibits a weak cooperative thermal transition. A significant increase in ANS fluorescence showed extensive solvent exposure of nonpolar cluster. Size exclusion chromatography (SEC) indicates a slight increase in the hydrodynamic size of acid-induced protein. These results suggest that asialofetuin at pH 1.8 represents the MG-like folding intermediate. Moreover, our results showed that glycosylation might play a role in stabilization of secondary structure during acid and/or thermal denaturation.
authors.affiliation: Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh 202002, India.
authors.forename: Farah
authors.initials: F
authors.lastname: Naseem
authors.forename: Basir
authors.initials: B
authors.lastname: Ahmad
authors.forename: Mohd Tashfeen
authors.initials: MT
authors.lastname: Ashraf
authors.forename: Rizwan Hasan
authors.initials: RH
authors.lastname: Khan
mesh_terms: D000143:Acids / Q000737:chemistry; D020106:Acrylamide / Q000737:chemistry; D000818:Animals; D001212:Asialoglycoproteins / Q000737:chemistry*; D002417:Cattle; D002942:Circular Dichroism; D060748:Fetuins; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence; D013696:Temperature; D014364:Tryptophan / Q000737:chemistry; D000509:alpha-Fetoproteins / Q000737:chemistry*
chemical_list: D000143:Acids; D001212:Asialoglycoproteins; D060748:Fetuins; D000509:alpha-Fetoproteins; C017029:asialofetuin; D020106:Acrylamide; D014364:Tryptophan