Cloning, preparation and preliminary crystallographic studies of penicillin V acylase autoproteolytic processing mutants.
The crystallization of three catalytically inactive mutants of penicillin V acylase (PVA) from Bacillus sphaericus in precursor and processed forms is reported. The mutant proteins crystallize in different primitive monoclinic space groups that are distinct from the crystal forms for the native enzyme. Directed mutants and clone constructs were designed to study the post-translational autoproteolytic processing of PVA. The catalytically inactive mutants will provide three-dimensional structures of precursor PVA forms, plus open a route to the study of enzyme-substrate complexes for this industrially important enzyme.
Chandra
P Manish
PM
Division of Biochemical Sciences, National Chemical Laboratory, Pune 411008, India.
Brannigan
James A
JA
Prabhune
Asmita
A
Pundle
Archana
A
Turkenburg
Johan P
JP
Dodson
G Guy
GG
Suresh
C G
CG
D019943:Amino Acid Substitution; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D003460:Crystallization; D004926:Escherichia coli / Q000201:enzymology; D016297:Mutagenesis, Site-Directed; D010399:Penicillin Amidase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D010399:Penicillin Amidase