C-Terminal Propeptide of BKA has a Protease Sensitive Structure Without any Inhibitory Effect on BKA

Document Type: Article

Authors

1 Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran

2 Nanobiotechnology Research Center, Baqiyatallah University of Medical Science, Tehran, Iran

Abstract

In our previous study, we compared the two α-amylase enzymes from Bacillus sp.KR8104, BKA∆(N44) and BKA∆(N44C193) which is the secreted form of it. The results indicated that the presence of 193 amino acids propeptide in the C-terminal of BKA∆(N44) changed its enzymatic parameters like an uncompetitive inhibitor in comparison to BKA∆(N44C193). In the present study, we cloned the DNA sequence of BKA∆(N44) which codes the 193 amino acids propeptide in its C-terminal and the effect of this fragment as an inhibitor on BKA∆(N44C193) was investigated. We also studied the possible foldase activity of the propeptide in BKA∆(N44C193). Protease sensitivity of C-terminal 193 amino acid propeptide, BKA∆(N44) and BKA∆(N44C193) was compared in order to explain why  BKA∆(N44C193) is the only secreted form of α-amylase in the culture medium of Bacillus sp.KR8104. Circular dichroism indicated that the secondary structure of the C-terminal is mostly beta sheeted. At the end we proposed a possible regulatory role for the C-terminal propeptide of BKA.

Graphical Abstract

C-Terminal Propeptide of BKA has a Protease Sensitive Structure Without any Inhibitory Effect on BKA

Keywords

Main Subjects


 

[1]           P. Bernfeld, Methods. Enzymol, Academic Press, 1955.

[2]           M.M. Bradford, Anal. Biochem. 72 (1976) 248.

[3]           Y.   Chen,   M. Inouye,  Curr.  Opin.  Struc.  Biol.  18 (2008) 765.

[4]           M. Desvaux, N.J.  Parham, I. R. Henderson, Res. Microbiol.155 (2004)53.

[5]           M. El Khattabi, P.V.  Gelder, W. Bitter, J. Tommassen, J. Biol. Chem. 275 (2000) 26885.

[6]           G. Feller, S.M. D’Amico, A. Benotmane, F. Joly, J. Van Beeumen, C. Gerday, J. Biol. Chem. 273(1998) 12109.

[7]           Z. Fujimoto, K. Takase, N. Doui, M. Momma, T. Matsumoto, H. Mizuno, J. Mol. Biol. 277(1998) 393.

[8]           I.R. Henderson, R. Cappello, J.P. Nataro, Trends. Microbiol.8(2000) 529.

[9]           I.R. Henderson, F. Navarro-Garcia, J.P. Nataro, Trends. Microbiol. 6 (1998) 370.

[10]       I.R. Henderson, F. Navarro-Garcia, M. Desvaux, R.C. Fernandez, D. Ala'Aldeen,Microbiol. Mol. Biol. Rev.68 (2004) 692.

[11]       M. Kagawa, Z. Fujimoto, M. Momma, K. Takase, H. Mizuno, J. Bacteriol.185 (2003) 6981.

[12]       U.K. Laemmli, Nature227 (1970) 680.

[13]       L.L. Lin, W.H. Hsu, W.S. Chu, J. Appl. Microbiol. 82 (1997)325.

[14]       M. Machius, G.  Wiegand,  R. Huber,  J.  Mol.  Biol.246(1995) 545.

[15]       K. Mclver, E. Kessler, J. Olson, D. Ohman. Mol. Microbiol. 18 (1995) 877.

[16]       T. Nakada, M. Kubota, S. Sakai, Y. Tsujisaka,Agric. Biol. Chem. 54 (1990)737.

[17]       K. Ohdan, T. Kuriki, H. Kaneco, J. Shimada, T. Takada, Z. Fujimoto, H. Mizuno, S. Okada,Appl. Environ. Microbiol. 65 (1999) 4652.

[18]       A. Salimi, F. Yousefi, M. Ghollasi, S. Daneshjou, H. Tavoli, S. Ghobadi, K. Khajeh, J. Microbiol. Biotechnol.22 (2012) 1077.

[19]       Y. Sakakibara, K. Tsutsumi, K. Nakamura, K. Yamane, J. Biochem.175 (1993)4203.

[20]       H. Sasamoto, K. Nakazawa, K. Tsutsumi, K. Takase, K .Yamane, J. Biochem.106 (1989) 376.

[21]       M. Søgaard, F.L. Olsen, B. Svensson, Proc. Natl. Acad. Sci. 88 (1991)8140.

[22]       K. Takase, H. Mizuno, K. Yamane, J. biol. Chem. 263 (1988)11548.

[23]       H. Tjalsma, A. Bolhuis, J.D.H. Jongbloed, S. Bron, J.M. van Dijl, Microbiol. Mol. Biol. Rev. 64 (2000)515.

[24]       C.M. Wilson, Methods Enzmol.91(1983) 236.