Tertiary Structure Formation at Specific Tryptophan Side Chains in the Refolding of Human Carbonic Anhydrase II

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• 作者：Per Jonasson ; G&ouml ; ran Aronsson ; Uno Carlsson ; and Bengt-Harald Jonsson
• 刊名：Biochemistry
• 出版年：1997
• 出版时间：April 29, 1997
• 年：1997
• 卷：36
• 期：17
• 页码：5142 - 5148
• 全文大小：268K
• 年卷期：v.36,no.17(April 29, 1997)
• ISSN：1520-4995

文摘

The refolding reaction of human carbonic anhydrase II has beencharacterized by use of sevenvariants in which tryptophan residues have been replaced by Phe or Cys,in each case giving proteinswith six tryptophans. Intrinsic tryptophan fluorescence was usedto monitor the refolding in the 2 ms-60 s time range, and kinetic traces showing the contributions from eachparticular tryptophan were obtainedby calculation of differences between the wild-type protein and thevariants. Earlier assignment[Mårtensson, L.-G., Jonasson, P., Freskgård, P.-O.,Svensson, M., Carlsson, U., & Jonsson, B.-H. (1995)Biochemistry 34, 1011-1021] of specific fluorescenceproperties to each tryptophan, especially regardingenergy transfer and intrinsic fluorescence quenching, has made itpossible to use the kinetic data to describethe formation of tertiary structure at defined tryptophan residues.In summary, it was found that tertiarystructure is formed earlier at those tryptophans that are associatedwith the central core of -strands thanat tryptophan residues in the N-terminal minidomain.