Cloning, expression, purification and crystallization as well as X-ray fluorescence and preliminary X-ray diffraction analyses of human ADP-ribosylhydrolase 1.
Human ADP-ribosylhydrolase 1 (hARH1, ADPRH) cleaves the glycosidic bond of ADP-ribose attached to an Arg residue of a protein. hARH1 has been cloned, expressed heterologously in Escherichia coli, purified and crystallized in complex with K(+) and ADP. The orthorhombic crystals contained one monomer per asymmetric unit, exhibited a solvent content of 43% and diffracted X-rays to a resolution of 1.9 A. A prerequisite for obtaining well diffracting crystals was the performance of X-ray fluorescence analysis on poorly diffracting apo hARH1 crystals, which revealed the presence of trace amounts of K(+) in the crystal. Adding K-ADP to the crystallization cocktail then resulted in a crystal of different morphology and with dramatically improved diffraction properties.
Kernstock
Stefan
S
Institut für Immunologie, Universitätsklinikum Eppendorf, Hamburg, Germany.
Koch-Nolte
Friedrich
F
Mueller-Dieckmann
Jochen
J
Weiss
Manfred S
MS
Mueller-Dieckmann
Christoph
C
D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004798:Enzymes / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005453:Fluorescence; D015870:Gene Expression; D009699:N-Glycosyl Hydrolases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014965:X-Rays
D004798:Enzymes; D009699:N-Glycosyl Hydrolases; C046422:ADP-ribosylarginine hydrolase