Salt-induced folding of alkaline denatured creatine kinase under high pH conditions

Hai Peng Yang, Haining Zhong, Sen Li, Hai Meng Zhou

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The conformational changes of creatine kinase during alkaline unfolding anti salt-induced folding at high pH have been followed by fluorescence emission and circular dichroism spectra. The results obtained show that at low ionic strength, with increasing pH value, creatine kinase denatured gradually to reach the ultimate unfolded conformation. in the vicinity of pH 12.7. With the increase of pH from 9.0 to 12.7, tile fluorescence emission maximum red shifted from 337 to 355 nm, indicating complete exposure of the buried tryptophan residues to the solvent. The far-UV CD spectra show that even at pH 12.7, the apparently fully denatured enzyme retains a great part of ordered secondary structure. At pH 12.7 by adding the salt, the relatively unfolded state of denatured enzyme changes into a compact conformational state by hydrophobic collapsing. Folded state induced by salt bound ANS strongly, indicating the existence of increased hydrophobic surface. The above results suggest that the salt-induced folded state at high pH may be the folded intermediate which exists in the general protein folding, and the larger residual ordered secondary structure might become folded being point on the salt-induced folding.

Original languageEnglish (US)
Pages (from-to)257-267
Number of pages11
JournalBiochemistry and Molecular Biology International
Volume41
Issue number2
StatePublished - Feb 1997
Externally publishedYes

Fingerprint

Creatine Kinase
Salts
Fluorescence
Protein folding
Enzymes
Tile
Ionic strength
Tryptophan
Conformations
Protein Folding
Circular Dichroism
Osmolar Concentration

Keywords

  • Alkaline denaturation
  • Creatine kinase
  • Molten globule state
  • Refolding

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Molecular Biology

Cite this

Salt-induced folding of alkaline denatured creatine kinase under high pH conditions. / Yang, Hai Peng; Zhong, Haining; Li, Sen; Zhou, Hai Meng.

In: Biochemistry and Molecular Biology International, Vol. 41, No. 2, 02.1997, p. 257-267.

Research output: Contribution to journalArticle

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N2 - The conformational changes of creatine kinase during alkaline unfolding anti salt-induced folding at high pH have been followed by fluorescence emission and circular dichroism spectra. The results obtained show that at low ionic strength, with increasing pH value, creatine kinase denatured gradually to reach the ultimate unfolded conformation. in the vicinity of pH 12.7. With the increase of pH from 9.0 to 12.7, tile fluorescence emission maximum red shifted from 337 to 355 nm, indicating complete exposure of the buried tryptophan residues to the solvent. The far-UV CD spectra show that even at pH 12.7, the apparently fully denatured enzyme retains a great part of ordered secondary structure. At pH 12.7 by adding the salt, the relatively unfolded state of denatured enzyme changes into a compact conformational state by hydrophobic collapsing. Folded state induced by salt bound ANS strongly, indicating the existence of increased hydrophobic surface. The above results suggest that the salt-induced folded state at high pH may be the folded intermediate which exists in the general protein folding, and the larger residual ordered secondary structure might become folded being point on the salt-induced folding.

AB - The conformational changes of creatine kinase during alkaline unfolding anti salt-induced folding at high pH have been followed by fluorescence emission and circular dichroism spectra. The results obtained show that at low ionic strength, with increasing pH value, creatine kinase denatured gradually to reach the ultimate unfolded conformation. in the vicinity of pH 12.7. With the increase of pH from 9.0 to 12.7, tile fluorescence emission maximum red shifted from 337 to 355 nm, indicating complete exposure of the buried tryptophan residues to the solvent. The far-UV CD spectra show that even at pH 12.7, the apparently fully denatured enzyme retains a great part of ordered secondary structure. At pH 12.7 by adding the salt, the relatively unfolded state of denatured enzyme changes into a compact conformational state by hydrophobic collapsing. Folded state induced by salt bound ANS strongly, indicating the existence of increased hydrophobic surface. The above results suggest that the salt-induced folded state at high pH may be the folded intermediate which exists in the general protein folding, and the larger residual ordered secondary structure might become folded being point on the salt-induced folding.

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