Crystallization and preliminary X-ray diffraction studies of AsaP1_E294A and AsaP1_E294Q, two inactive mutants of the toxic zinc metallopeptidase AsaP1 from Aeromonas salmonicida subsp. achromogenes.
Two mutants of the toxic extracellular zinc endopeptidase AsaP1 (AsaP1_E294Q and AsaP1_E294A) of Aeromonas salmonicida subsp. achromogenes were expressed in Escherichia coli and crystallized by the vapour-diffusion method. Crystals were obtained using several precipitants and different protein concentrations. Protein crystals were found in a monoclinic (C2) as well as an orthorhombic (P2(1)2(1)2(1)) space group. The crystals belonging to the monoclinic space group C2 had unit-cell parameters a = 103.4, b = 70.9, c = 54.9 A, beta = 109.3 degrees for AsaP1_E294A, and a = 98.5, b = 74.5, c = 54.7 A, beta = 112.4 degrees for AsaP1_E294Q. The unit-cell parameters of the orthorhombic crystal obtained for AsaP1_E294A were a = 57.9, b = 60.2, c = 183.6 A. The crystals of the two different mutants diffracted X-rays beyond 2.0 A resolution.
Bogdanović
Xenia
X
Institut für Biochemie, Universität Greifswald, D-17489 Greifswald, Germany.
Singh
Rajesh Kumar
RK
Hentschke
Johanna
J
Gudmundsdóttir
Bjarnheidur K
BK
Hinrichs
Winfried
W
D048409:Aeromonas salmonicida / Q000201:enzymology*; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D045726:Metalloproteases / Q000737:chemistry*; D050505:Mutant Proteins / Q000737:chemistry*
D001426:Bacterial Proteins; D050505:Mutant Proteins; D045726:Metalloproteases