Structure of the imipenem-hydrolyzing class A beta-lactamase SME-1 from Serratia marcescens.
The structure of the beta-lactamase SME-1 from Serratia marcescens, a class A enzyme characterized by its significant activity against imipenem, has been determined to 2.13 A resolution. The overall structure of SME-1 is similar to that of other class A beta-lactamases. In the active-site cavity, most of the residues found in SME-1 are conserved among class A beta-lactamases, except at positions 104, 105 and 237, where a tyrosine, a histidine and a serine are found, respectively, and at position 238, which is occupied by a cysteine forming a disulfide bridge with the other cysteine residue located at position 69. The crucial role played by this disulfide bridge in SME-1 was confirmed by site-directed mutagenesis of Cys69 to Ala, which resulted in a mutant unable to confer resistance to imipenem and all other beta-lactam antibiotics tested. Another striking structural feature found in SME-1 was the short distance separating the side chains of the active serine residue at position 70 and the strictly conserved glutamate at position 166, which is up to 1.4 A shorter in SME-1 compared with other class A beta-lactamases. Consequently, the SME-1 structure cannot accommodate the essential catalytic water molecule found between Ser70 and Glu166 in the other class A beta-lactamases described so far, suggesting that a significant conformational change may be necessary in SME-1 to properly position the hydrolytic water molecule involved in the hydrolysis of the acyl-enzyme intermediate.
Sougakoff
Wladimir
W
Laboratoire de Recherche Moléculaire sur les Antibiotiques (LRMA), INSERM EMI0004, Faculté de Médecine Pitié-Salpêtrière, Université Pierre et Marie Curie, 75634 Paris CEDEX 13, France.
L'Hermite
Guillaume
G
Pernot
Lucile
L
Naas
Thierry
T
Guillet
Valérie
V
Nordmann
Patrice
P
Jarlier
Vincent
V
Delettré
Jean
J
D000409:Alanine / Q000235:genetics; D000595:Amino Acid Sequence; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000235:genetics; D006868:Hydrolysis; D015378:Imipenem / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D012706:Serratia marcescens / Q000201:enzymology*; D001618:beta-Lactamases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D015378:Imipenem; C094219:beta-lactamase Sme-1; D001618:beta-Lactamases; D003545:Cysteine; D000409:Alanine