2015
DOI: 10.1107/s2053230x15000424
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Crystallization and preliminary X-ray crystallographic studies of dipeptidyl peptidase 11 fromPorphyromonas gingivalis

Abstract: Dipeptidyl peptidase 11 from Porphyromonas gingivalis (PgDPP11) preferentially cleaves substrate peptides with Asp and Glu at the P1 position [NH2-P2-P1(Asp/Glu)-P1'-P2'...]. For crystallographic studies, PgDPP11 was overproduced in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. X-ray diffraction data to 1.82 Å resolution were collected from an orthorhombic crystal form belonging to space group C2221, with unit-cell parameters a = 99.33, b = 103.60, c = 177.33 Å. St… Show more

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“…PgDPP11 was expressed and purified as described elsewhere 48 . A synthetic gene coding for full-length PgDPP11 (residues 1-720), codon-optimised for expression in E. coli , was purchased from Genscript (Piscataway, USA) and cloned into the pET22b expression plasmid (Merck, Darmstadt, Germany).…”
Section: Methodsmentioning
confidence: 99%
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“…PgDPP11 was expressed and purified as described elsewhere 48 . A synthetic gene coding for full-length PgDPP11 (residues 1-720), codon-optimised for expression in E. coli , was purchased from Genscript (Piscataway, USA) and cloned into the pET22b expression plasmid (Merck, Darmstadt, Germany).…”
Section: Methodsmentioning
confidence: 99%
“…To obtain peptide-free PgDPP11 crystals 48 , the samples were crystallised using the hanging-drop method, in which 1 μl of protein solution (10 mg/ml PgDPP11 in 80 mM Tris-HCl, pH 8.5) was mixed with the same volume of reservoir solution (20% (v/v) glycerol, 16% (m/v) PEG8000 and 0.16 M tri-potassium citrate) and incubated at 293 K. The drops were suspended over 200 μl of reservoir solution in 48-well plates. Peptide-free crystals were also obtained using a counter-diffusion crystallisation method 49 under a microgravity environment in the Japanese Experimental Module “Kibo” at the International Space Station (ISS) 47 .…”
Section: Methodsmentioning
confidence: 99%