Tatu, Utpal and Murthy, SK and Vithayathil, PJ (1990) Role of a disulfide cross-link in the conformational stability of a thermostable xylanase. In: Journal of Protein Chemistry, 9 (5). pp. 641-646.
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The role of a S-S cross-link in the conformational stability of xylanase fromHumicola lanuginosa has been investigated using CD, UV absorption spectroscopy, and RIA displacement studies. Our studies show that reduction and carboxymethylation of the S-S cross-link in xylanase results in a gross conformational perturbation of the protein. The secondary structure analysis of the CD spectra indicates that the xylanase with an intact S-S contains 66% $\beta$-sheet structure and remaining random coil. Cleavage of the S-S bond results in a loss of 25% $\beta$-sheet structure. Thermal denaturation studies using CD spectroscopy and pH-dependent tyrosine ionization studies using UV spectroscopy show that the presence of disulfide cross-link offers resistance against unfolding by extremes of temperature and pH. Further, we demonstrate that the heat-induced changes in xylanase with intact S-S bond are almost totally reversible, while those in the S-S cleaved enzyme fail to show any significant reversal. Our studies support the present theory that S-S cross-links exert their stabilizing effect in proteins by destabilizing the unfolded state of the protein and forcing it back to a more folded state.
|Item Type:||Journal Article|
|Additional Information:||Copyright of this article belongs to Plenum Publishing Corporafioa.|
|Keywords:||CD;disulfide bond;protein stability;RIA;secondary structure;tyrosine ionization;xylanase|
|Department/Centre:||Division of Biological Sciences > Biochemistry|
|Date Deposited:||10 Dec 2007|
|Last Modified:||19 Sep 2010 04:41|
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