Pak. J. Bot., 43(4): 2059-2065, 2011. | Back to Contents | ||||
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REQUIREMENT OF
PRO-PEPTIDE IN PROPER FOLDING OF SUBTILISIN-LIKE SERINE
PROTEASE TK0076
Abstract:
Subtilisin-like
proteases are characterized by a catalytic triad of the three amino
acids Asp His and Ser. Tk0076 is a subtilisin-like serine protease
originated from Thermococcus kodakaraensis. Regions corresponding
to signal-peptide, pro-peptide and the mature protein were predicted by
homology modeling. Homology comparison revealed that Asp215,
His247 and Ser424 constitute the catalytic triad
of the protein. Gene encoding Tk0076 was cloned and expressed in
Escherichia coli. The protein was produced in the soluble form when
the gene contained the sequence corresponding to the pro-peptide whereas
it was produced in the insoluble form without the sequence corresponding
to pro-peptide under the same expression system. Attempts to refold the
protein properly in the absence of pro-peptide were unsuccessful
indicating that pro-peptide is essential for proper folding of Tk0076.
School of Biological
Sciences, University of the Punjab, Quaid-e-Azam Campus, Lahore 54590,
Pakistan |
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