[en] The peptidoglycan glycosyltransferase (GT) module of class A penicillin-binding proteins (PBPs) and monofunctional GTs catalyze glycan chain elongation of the bacterial cell wall. These enzymes belong to the GT51 family, are characterized by five conserved motifs, and have some fold similarity with the phage lambda lysozyme. In this work, we have systematically modified all the conserved amino acid residues of the GT module of Escherichia coli class A PBP1b by site-directed mutagenesis and determined their importance for the in vivo and in vitro activity and the thermostability of the protein. To get an insight into the GT active site of this paradigm enzyme, a model of PBP1b GT domain was constructed based on the available crystal structures (PDB codes 2OLV and 2OLU). The data show that in addition to the essential glutamate residues Glu233 of motif 1 and Glu290 of motif 3, the residues Phe237 and His240 of motif 1 and Gly264, Thr267, Gln271, and Lys274 of motif 2, all located in the catalytic cavity of the GT domain, are essential for the in vitro enzymatic activity of the PBP1b and for its in vivo functioning. Thus, the first three conserved motifs contain most of the residues that are required for the GT activity of the PBP1b. The residues Asp234, Phe237, His240, Thr267, and Gln271 are proposed to maintain the structure of the active site and the positioning of the catalytic Glu233.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Terrak, Mohammed ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Sauvage, Eric ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Derouaux, Adeline ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Dehareng, Dominique ; Université de Liège - ULiège > Centre d'ingénierie des protéines
scite shows how a scientific paper has been cited by providing the context of the citation, a classification describing whether it supports, mentions, or contrasts the cited claim, and a label indicating in which section the citation was made.
Bibliography
van Heijenoort, J. (2001) Glycobiology 11, 25R-36R
Vollmer, W., Blanot, D., and de Pedro, M. A. (2008) FEMS Microbiol. Rev. 32, 149-167
Vollmer, W., and Höltje, J. V. (2001) Curr. Opin. Microbiol. 4, 625-633
Nanninga, N. (1991) Mol. Microbiol. 5, 791-795
Goehring, N. W., and Beckwith, J. (2005) Curr. Biol. 15, R514-526
Höltje, J. V. (1998) Microbiol. Mol. Biol. Rev. 62, 181-203
den Blaauwen, T., de Pedro, M. A., Nguyen-Distèche, M., and Ayala, J. A. (2008) FEMS Microbiol. Rev. 32, 321-344
Goffin, C., and Ghuysen, J. M. (1998) Microbiol. Mol. Biol. Rev. 62, 1079-1093
Sauvage, E., Kerff, F., Terrak, M., Ayala, J. A., and Charlier, P. (2008) FEMS Microbiol. Rev. 32, 234-258
Marrec-Fairley, M., Piette, A., Gallet, X., Brasseur, R., Hara, H., Fraipont, C., Ghuysen, J. M., and Nguyen-Distèche, M. (2000) Mol. Microbiol. 37, 1019-1031
Spratt, B. G., Zhou, J., Taylor, M., and Merrick, M. J. (1996) Mol. Microbiol. 19, 639-640
Lovering, A. L., de Castro, L. H., Lim, D., and Strynadka, N. C. (2007) Science 315, 1402-1405
Yuan, Y., Barrett, D., Zhang, Y., Kahne, D., Sliz, P., and Walker, S. (2007) Proc. Natl. Acad. Sci. U. S. A. 104, 5348-5353
Perlstein, D. L., Zhang, Y., Wang, T. S., Kahne, D. E., and Walker, S. (2007) J. Am. Chem. Soc. 129, 12674-12675
Barrett, D., Wang, T. S., Yuan, Y., Zhang, Y., Kahne, D., and Walker, S. (2007) J. Biol. Chem. 282, 31964-31971
Schwartz, B., Markwalder, J. A., Seitz, S. P., Wang, Y., and Stein, R. L. (2002) Biochemistry 41, 12552-12561
Terrak, M., Ghosh, T. K., van Heijenoort, J., Van Beeumen, J., Lampilas, M., Aszodi, J., Ayala, J. A., Ghuysen, J. M., and Nguyen-Distèche, M. (1999) Mol. Microbiol. 34, 350-364
Barrett, D., Leimkuhler, C., Chen, L., Walker, D., Kahne, D., and Walker, S. (2005) J. Bacteriol. 187, 2215-2217
Zawadzka-Skomial, J., Markiewicz, Z., Nguyen-Distèche, M., Devreese, B., Frère, J. M., and Terrak, M. (2006) J. Bacteriol. 188, 1875-1881
Terrak, M., and Nguyen-Distèche, M. (2006) J. Bacteriol. 188, 2528-2532
Tamaki, S., Nakajima, S., and Matsuhashi, M. (1977) Proc. Natl. Acad. Sci. U. S. A. 74, 5472-5476
Suzuki, H., Nishimura, Y., and Hirota, Y. (1978) Proc. Natl. Acad. Sci. U. S. A. 75, 664-668
Suzuki, H., van Heijenoort, Y., Tamura, T., Mizoguchi, J., Hirota, Y., and van Heijenoort, J. (1980) FEBS Lett. 110, 245-249
Bertsche, U., Breukink, E., Kast, T., and Vollmer, W. (2005) J. Biol. Chem. 280, 38096-38101
Bertsche, U., Kast, T., Wolf, B., Fraipont, C., Aarsman, M. E., Kannenberg, K., von Rechenberg, M., Nguyen-Distèche, M., den Blaauwen, T., Höltje, J. V., and Vollmer, W. (2006) Mol. Microbiol. 61, 675-690
Vollmer, W., von Rechenberg, M., and Höltje, J. V. (1999) J. Biol. Chem. 274, 6726-6734
Müller, P., Ewers, C., Bertsche, U., Anstett, M., Kallis, T., Breukink, E., Fraipont, C., Terrak, M., Nguyen-Distèche, M., and Vollmer, W. (2007) J. Biol. Chem. 282, 36394-36402
Barrett, D. S., Chen, L., Litterman, N. K., and Walker, S. (2004) Biochemistry 43, 12375-12381
Coutinho, P. M., Deleury, E., Davies, G. J., and Henrissat, B. (2003) J. Mol. Biol. 328, 307-317
Hara, H., and Suzuki, H. (1984) FEBS Lett. 168, 155-160
van Heijenoort, Y., Gomez, M., Derrien, M., Ayala, J., and van Heijenoort, J. (1992) J. Bacteriol. 174, 3549-3557
Lakaye, B., Damblon, C., Jamin, M., Galleni, M., Lepage, S., Joris, B., Marchand-Brynaert, J., Frydrych, C., and Frère, J. M. (1994) Biochem. J. 300, 141-145
Similar publications
Sorry the service is unavailable at the moment. Please try again later.
This website uses cookies to improve user experience. Read more
Save & Close
Accept all
Decline all
Show detailsHide details
Cookie declaration
About cookies
Strictly necessary
Performance
Strictly necessary cookies allow core website functionality such as user login and account management. The website cannot be used properly without strictly necessary cookies.
This cookie is used by Cookie-Script.com service to remember visitor cookie consent preferences. It is necessary for Cookie-Script.com cookie banner to work properly.
Performance cookies are used to see how visitors use the website, eg. analytics cookies. Those cookies cannot be used to directly identify a certain visitor.
Used to store the attribution information, the referrer initially used to visit the website
Cookies are small text files that are placed on your computer by websites that you visit. Websites use cookies to help users navigate efficiently and perform certain functions. Cookies that are required for the website to operate properly are allowed to be set without your permission. All other cookies need to be approved before they can be set in the browser.
You can change your consent to cookie usage at any time on our Privacy Policy page.