Crystallization and preliminary X-ray diffraction analysis of the extracellular domain of the cell surface antigen CD38 complexed with ganglioside.
The cell surface antigen CD38 is a multifunctional ectoenzyme that acts as an NAD(+) glycohydrolase, an ADP-ribosyl cyclase, and also a cyclic ADP-ribose hydrolase. The extracellular catalytic domain of CD38 was expressed as a fusion protein with maltose-binding protein, and was crystallized in the complex with a ganglioside, G(T1b), one of the possible physiological inhibitors of this ectoenzyme. Two different crystal forms were obtained using the hanging-drop vapor diffusion method with PEG 10,000 as the precipitant. One form diffracted up to 2.4 A resolution with synchrotron radiation at 100 K, but suffered serious X-ray damage. It belongs to the space group P2(1)2(1)2(1) with unit-cell parameters of a = 47.9, b = 94.9, c = 125.2 A. The other form is a thin plate, but the data sets were successfully collected up to 2.4 A resolution by use of synchrotron radiation at 100 K. The crystals belong to the space group P2(1) with unit-cell parameters of a = 57.4, b = 51.2, c = 101.1 A, and beta = 97.9 degrees, and contain one molecule per asymmetric unit with a VM value of 2.05 A(3)/Da.
Kukimoto
M
M
Cellular Signaling Laboratory, Frontier Research Program, RIKEN (The Institute of Physical and Chemical Research), Hirosawa, Wako-shi, Saitama, 351-0198, Japan.
Nureki
O
O
Shirouzu
M
M
Katada
T
T
Hirabayashi
Y
Y
Sugiya
H
H
Furuyama
S
S
Yokoyama
S
S
Hara-Yokoyama
M
M
D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003460:Crystallization; D005732:Gangliosides / Q000737:chemistry* / Q000378:metabolism*; D058619:Maltose-Binding Proteins; D009244:NAD+ Nucleosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014961:X-Ray Diffraction
D015703:Antigens, CD; D000943:Antigens, Differentiation; D002352:Carrier Proteins; D005732:Gangliosides; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins; C016481:trisialoganglioside GT1; D036541:ADP-ribosyl Cyclase; D009244:NAD+ Nucleosidase; D051997:ADP-ribosyl Cyclase 1