Domain organization of penicillin-binding protein 5 from Escherichia coli analysed by C-terminal truncation.

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Citation

van der Linden MP, de Haan L, Keck W

Domain organization of penicillin-binding protein 5 from Escherichia coli analysed by C-terminal truncation.

Biochem J. 1993 Jan 15;289 ( Pt 2):593-8.

PubMed ID
8424800 [ View in PubMed
]
Abstract

The structural organization of penicillin-binding protein (PBP) 5 was investigated by C-terminal truncation. Compared with other low-M(r) penicillin-interacting proteins, PBP5 carries a C-terminal extension of about 100 amino acids. The sites for introduction of stop codons were chosen on the basis of the established three-dimensional structure of the Streptomyces albus G beta-lactamase [Dideberg, Charlier, Wery, Dehottay, Dusart, Erpicum, Frere and Ghuysen (1987) Biochem. J. 245, 911-913] and comparative hydrophobic cluster analysis [Gaboriaud, Bissery, Bencheritt and Mornon (1987) FEBS Lett. 224, 149-155]. Two stop codons were introduced at positions Ile-354 or Val-348 to construct an optimized soluble form of PBP5 for crystallization purposes. The newly constructed soluble and enzymically active form (PBP5s353) was isolated by dye-affinity chromatography and gave rise to small crystals. Another two stop codons were introduced at positions Arg-261 or Ala-276 to determine the minimal enzymically active 'core protein'. The truncated form (PBP5s275), missing the entire C-terminal extension, showed unaltered penicillin-binding characteristics and a catalytic-centre activity 40% that of PBP5s353 + 9 using bisacetyl-L-Lys-D-Ala-D-Ala as substrate. This protein, however was more susceptible to proteolytic degradation, which might indicate a role of the C-terminal portion in stabilizing the protein.

DrugBank Data that Cites this Article

Polypeptides
NameUniProt ID
D-alanyl-D-alanine carboxypeptidase DacAP0AEB2Details