The crystal structure of the <i>β</i>-lactamase of <i>Streptomyces albus</i> G at 0.3 nm resolution

Dideberg, Otto; Charlier, Paulette; Wery, J. P.; Dehottay, Philippe; Dusart, Jean; Erpicum, T.; Frère, Jean‐Marie; Ghuysen, Jean‐Marie

doi:10.1042/bj2450911

Cited by 148 publications

(101 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…10,11,14,18 In the b-lactamase from Streptomyces albus G, an arginine extends from position 220 rather than from position 244. 19 In support of an analogous role for Arg220 in substrate binding and catalysis, an R220L mutation of the Streptomyces albus G b-lactamase greatly impairs the catalytic ability of this enzyme. 15 The Streptomyces albus G b-lactamase structure includes an additional positively charged residue at nearby position 274 (Fig.…”

Section: Resultsmentioning

confidence: 98%

Analysis of the plasticity of location of the Arg244 positive charge within the active site of the TEM‐1 β‐lactamase

2009

View full text Add to dashboard Cite

A large number of b-lactamases have emerged that are capable of conferring bacterial resistance to b-lactam antibiotics. Comparison of the structural and functional features of this family has refined understanding of the catalytic properties of these enzymes. An arginine residue present at position 244 in TEM-1 b-lactamase interacts with the carboxyl group common to penicillin and cephalosporin antibiotics and thereby stabilizes both the substrate and transition state complexes. A comparison of class A b-lactamase sequences reveals that arginine at position 244 is not conserved, although a positive charge at this structural location is conserved and is provided by an arginine at positions 220 or 276 for those enzymes lacking arginine at position 244. The plasticity of the location of positive charge in the b-lactamase active site was experimentally investigated by relocating the arginine at position 244 in TEM-1 b-lactamase to positions 220, 272, and 276 by site-directed mutagenesis. Kinetic analysis of the engineered b-lactamases revealed that removal of arginine 244 by alanine mutation reduced catalytic efficiency against all substrates tested and restoration of an arginine at positions 272 or 276 partially suppresses the catalytic defect of the Arg244Ala substitution. These results suggest an evolutionary mechanism for the observed divergence of the position of positive charge in the active site of class A b-lactamases.

show abstract

Section: Resultsmentioning

confidence: 98%

Analysis of the plasticity of location of the Arg244 positive charge within the active site of the TEM‐1 β‐lactamase

2009

View full text Add to dashboard Cite

show abstract

“…Eight X-ray structures of apo-enzymes, TEM-1 and TOHO-1 from Escherichia coli (16,17), PC1 from Staphylococcus aureus (18), SHV from Klebsiella pneumoniae (19), NMC-A from Enterobacter cloacae (20), MFO from Mycobacterium fortuitum (PDB entry: 1MFO), BLIC from Bacillus licheniformis (21), and SAG from Streptomyces albus G (22) have been solved. These enzymes display a very similar fold and the detailed comparisons of the structures were helpful in relating some by guest on May 9, 2018 http://www.jbc.org/ Downloaded from 5 significant differences in substrate profile to local structural features (23,24).…”

Section: Introductionmentioning

confidence: 99%

The High Resolution Crystal Structure for Class A β-Lactamase PER-1 Reveals the Bases for Its Increase in Breadth of Activity

Tranier¹,

Bouthors²,

Maveyraud³

et al. 2000

Journal of Biological Chemistry

View full text Add to dashboard Cite

SUMMARYThe treatment of infectious diseases by β-lactam antibiotics is continuously challenged by the emergence and dissemination of new β-lactamases. In most cases, the cephalosporinase activity of class A enzymes results from a few mutations in the TEM and SHV penicillinases. The PER-1 β-lactamase was characterized as a class A enzyme displaying a cephalosporinase activity. This activity was however insensitive to the mutations of residues known to be critical for providing extended substrate profiles to TEM and SHV. The X-ray structure of the protein, solved at 1.9 Å resolution, reveals that two of the most conserved features in class A β-lactamase are not present in this enzyme: the fold of the Ω-loop and the cis conformation of the peptide bond between residues 166 and 167. The new fold of the Ω-loop and the insertion of four residues at the edge of strand S3 generate a broad cavity that may easily accomodate the bulky substituents of cephalosporin substrates. The trans conformation of the 166-167 bond is related to the presence of an aspartic acid at position 136. Selection of class A enzymes based on the occurrence of both Asp136 and Asn179 identifies a subgroup of enzymes with high sequence homology.

show abstract

“…Further conserved boxes in PBPs are the SXN sequence and the triad KTG or KSG that are part of the active-site cavity as deduced from the known three-dimensional structures of several p-lactamases and the Streptomyces R61 low-MI PBP (Dideberg et al, 1987;Herzberg and Moult, 1987;Moews et al, 1990;Oefner et al, 1990). Although not related in amino acid sequence, the polypeptide folding of the R61 PBP and the class A plactamases is amazingly similar with the conserved boxes representing critical components of the active-site cavity.…”

mentioning

confidence: 99%

Penicillin‐binding protein 2x of Streptococcus pneumoniae

Laible

Keck

Mottl

et al. 1992

European Journal of Biochemistry

View full text Add to dashboard Cite

A 2.5-kb DNA fragment including the structural gene coding for the penicillin-binding protein 2x (PBP 2x) of Streptococcus pneumoniue has been cloned into the vector pJDC9 and expressed in Escherichia coli. Mapping of RNA polymerase binding sites by electron microscopy indicated that the pbpX promoter is well recognized by the E. coli enzyme. However, high-level expression occurred mainly under the control of the lac promoter upstream of the pJDC9 multiple cloning site. After induction with isopropyl P-d-thiogalactopyranoside, PBP 2x was expressed as one of the major cellular proteins. PBP 2x produced in E. coli corresponded to the pneumococcal PBP 2x in terms of electrophoretic mobility, fractionation with the cytoplasmic membrane, and penicillin-binding capacity. Deletion of 30 hydrophobic N-terminal amino acid residues at positions 19 -48 resulted in high-level expression of a cytoplasmic, soluble PBP 2x derivative (PBP 2x*) which still retained full p-lactam-binding activity. A two-step procedure involving dye affinity chromatography was established for obtaining large amounts of highly purified enzymatically active PBP 2x*.

show abstract

The crystal structure of the β-lactamase of Streptomyces albus G at 0.3 nm resolution

Cited by 148 publications

References 14 publications

Analysis of the plasticity of location of the Arg244 positive charge within the active site of the TEM‐1 β‐lactamase

Analysis of the plasticity of location of the Arg244 positive charge within the active site of the TEM‐1 β‐lactamase

The High Resolution Crystal Structure for Class A β-Lactamase PER-1 Reveals the Bases for Its Increase in Breadth of Activity

Penicillin‐binding protein 2x of Streptococcus pneumoniae

Contact Info

Product

Resources

About