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X-ray crystal structures of the S229A mutant and wild-type MurB in the presence of the substrate enolpyruvyl-UDP-N-acetylglucosamine at 1.8-A resolution.
Biochemistry. 1997 Jan 28; 36(4):806-11.B

Abstract

MurB catalyzes the second committed step in the synthesis of peptidoglycan, a key component of the bacterial cell wall. The crystal structures of both a S229A mutant and wild-type MurB in the presence of the substrate enolpyruvyl-UDP-N-acetylglucosamine were solved and refined at 1.8 A resolution. The single point mutation of residue 229 from serine to alanine eliminated a hydroxyl group which has previously been proposed to play a critical role as a proton donor during the second half-reaction of MurB, namely, reoxidation of FADH2 and reduction of the enolpyruvyl substrate. The mutation also resulted in the loss of the water molecule-hydrogen bonded to the serine hydroxyl in the wild-type structure changing the hydrogen-bonding network with in the active site. Comparison of the wild-type and S229A mutant structures confirms that the dramatic kinetic defect of an approximately 10(7)-fold decrease observed for the Ser 229 Ala mutant in the second half-reaction [Benson, T.E., Walsh, C.T., & Massey, V. (1997) Biochemistry 36, 796-805] is a direct result of the loss of the serine hydroxyl moiety rather than other nonspecific active-site changes or general structural defects.

Authors+Show Affiliations

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

9020778

Citation

Benson, T E., et al. "X-ray Crystal Structures of the S229A Mutant and Wild-type MurB in the Presence of the Substrate enolpyruvyl-UDP-N-acetylglucosamine at 1.8-A Resolution." Biochemistry, vol. 36, no. 4, 1997, pp. 806-11.
Benson TE, Walsh CT, Hogle JM. X-ray crystal structures of the S229A mutant and wild-type MurB in the presence of the substrate enolpyruvyl-UDP-N-acetylglucosamine at 1.8-A resolution. Biochemistry. 1997;36(4):806-11.
Benson, T. E., Walsh, C. T., & Hogle, J. M. (1997). X-ray crystal structures of the S229A mutant and wild-type MurB in the presence of the substrate enolpyruvyl-UDP-N-acetylglucosamine at 1.8-A resolution. Biochemistry, 36(4), 806-11.
Benson TE, Walsh CT, Hogle JM. X-ray Crystal Structures of the S229A Mutant and Wild-type MurB in the Presence of the Substrate enolpyruvyl-UDP-N-acetylglucosamine at 1.8-A Resolution. Biochemistry. 1997 Jan 28;36(4):806-11. PubMed PMID: 9020778.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - X-ray crystal structures of the S229A mutant and wild-type MurB in the presence of the substrate enolpyruvyl-UDP-N-acetylglucosamine at 1.8-A resolution. AU - Benson,T E, AU - Walsh,C T, AU - Hogle,J M, PY - 1997/1/28/pubmed PY - 1997/1/28/medline PY - 1997/1/28/entrez SP - 806 EP - 11 JF - Biochemistry JO - Biochemistry VL - 36 IS - 4 N2 - MurB catalyzes the second committed step in the synthesis of peptidoglycan, a key component of the bacterial cell wall. The crystal structures of both a S229A mutant and wild-type MurB in the presence of the substrate enolpyruvyl-UDP-N-acetylglucosamine were solved and refined at 1.8 A resolution. The single point mutation of residue 229 from serine to alanine eliminated a hydroxyl group which has previously been proposed to play a critical role as a proton donor during the second half-reaction of MurB, namely, reoxidation of FADH2 and reduction of the enolpyruvyl substrate. The mutation also resulted in the loss of the water molecule-hydrogen bonded to the serine hydroxyl in the wild-type structure changing the hydrogen-bonding network with in the active site. Comparison of the wild-type and S229A mutant structures confirms that the dramatic kinetic defect of an approximately 10(7)-fold decrease observed for the Ser 229 Ala mutant in the second half-reaction [Benson, T.E., Walsh, C.T., & Massey, V. (1997) Biochemistry 36, 796-805] is a direct result of the loss of the serine hydroxyl moiety rather than other nonspecific active-site changes or general structural defects. SN - 0006-2960 UR - https://www.unboundmedicine.com/medline/citation/9020778/X_ray_crystal_structures_of_the_S229A_mutant_and_wild_type_MurB_in_the_presence_of_the_substrate_enolpyruvyl_UDP_N_acetylglucosamine_at_1_8_A_resolution_ L2 - https://doi.org/10.1021/bi962221g DB - PRIME DP - Unbound Medicine ER -