Preliminary crystallographic studies of Y25F mutant of periplasmic Escherichia coli L-asparaginase

• Autorzy: Kozak M. , Jaskolski M. , Rohm K.H.
• Języki publikacji: EN

Abstrakty

EN

Periplasmic Escherichia coli L-asparaginase II with Y25F mutation in the active-site cavity has been obtained by recombinant techniques. The protein was crystallized in a new hexagonal form (P6522). Single crystals of this polymorph, suitable for X-ray diffraction, were obtained by vapor diffusion using 2-methyl-2,4-pentanediol as precipitant (pH 4.8). The crystals are characterized by a = 81.0, c = 341.1 A and diffract to 2.45 A resolution. The asymmetric unit contains two protein molecules arranged into an AB dimer. The physiologically relevant ABA'B' homotetramer is generated by the action of the crystallographic 2-fold axis along [1, -1, 0]. Kinetic studies show that the loss of the phenolic hydroxyl group at position 25 brought about by the replacement of Y with F strongly impairs kcat without significantly affecting Km.

Słowa kluczowe

EN

L-asparaginase; leukemia; protein crystallography; protein; mutagenesis; Y25F mutation; amidohydrolase; mutant; Y25F mutant

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2000
• Tom: 47
• Numer: 3
• Opis fizyczny: p.807-814,fig.

Twórcy

autor

Kozak M.

• A.Mickiewicz University, Umultowska 85, 61-614 Poznan, Poland

autor

Jaskolski M.

autor

Rohm K.H.

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