PMID:21327882
| Citation |
Haghani, K, Khajeh, K, Salmanian, AH, Ranjbar, B and Bakhtiyari, S (2011) Acid-induced formation of molten globule states in the wild type Escherichia coli 5-enolpyruvylshikimate 3-phosphate synthase and its three mutated forms: G96A, A183T and G96A/A183T. Protein J. 30:132-7 |
|---|---|
| Abstract |
Recent advances in protein chemistry have led to progress in the understanding of protein folding and properties of possible intermediates during the folding of proteins. The molten globule (MG) state, a major intermediate of protein folding, has a denatured state with native-like secondary structure. In the present work, the acid-induced unfolding of wild type Escherichia coli 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) and its three different variants (G96A, A183T and G96A/A183T) were studied by far- and near-UV circular dichroism (CD), intrinsic fluorescent emission spectroscopy and 1-anilino naphthalene-8-sulfonate (ANS) binding. At pH < 3.0, these EPSPS variants acquire partially folded state, which show the characteristics of the MG state, e.g., a drastic reduction of defined tertiary structure and almost no change in the secondary structure. ANS binding experiments show that hydrophobic surface of these variants is exposed to a greater extent in comparison to the native form, at acidic pH. Wild type, G96A, A183T and G96A/A183T acquire MG states at pH 2.0, 1.5, 3.0 and 3.0, respectively, which show that pH stability of MG state of G96A has increased in comparison to wild type; and pH stability of MG states of two other mutants is lower than that of the wild type. The results suggest that there is a direct relationship between stability of protein and pH stability of its folding intermediates. |
| Links |
PubMed Online version:10.1007/s10930-011-9308-2 |
| Keywords |
3-Phosphoshikimate 1-Carboxyvinyltransferase/chemistry; 3-Phosphoshikimate 1-Carboxyvinyltransferase/genetics; Acids/chemistry; Anilino Naphthalenesulfonates/chemistry; Circular Dichroism; Enzyme Stability; Escherichia coli/enzymology; Hydrogen-Ion Concentration; Mutation; Protein Binding; Protein Folding; Protein Structure, Secondary; Spectrometry, Fluorescence; Ultraviolet Rays |
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