Expression, purification, crystallization and preliminary X-ray analysis of the N-terminal domain of GNBP3 from Drosophila melanogaster.
Gram-negative bacteria-binding protein 3 (GNBP3) is a pattern-recognition receptor which contributes to the defensive response against fungal infection in Drosophila. The protein consists of an N-terminal domain, which is considered to recognize beta-glucans from the fungal cell wall, and a C-terminal domain, which is homologous to bacterial glucanases but devoid of activity. The N-terminal domain of GNBP3 (GNBP3-Nter) was successfully purified after expression in Drosophila S2 cells. Diffraction-quality crystals were produced by the hanging-drop vapour-diffusion method using PEG 2000 and PEG 8000 as precipitants. Preliminary X-ray diffraction analysis revealed that the GNBP3-Nter crystals belonged to the monoclinic space group C2, with unit-cell parameters a = 134.79, b = 30.55, c = 51.73 A, beta = 107.4 degrees, and diffracted to 1.7 A using synchrotron radiation. The asymmetric unit is expected to contain two copies of GNBP3-Nter. Heavy-atom derivative data were collected and a samarium derivative showed one high-occupancy site per molecule.
Mishima
Yumiko
Y
Centre de Biophysique Moléculaire, UPR 4301 CNRS, Orléans, France.
Coste
Franck
F
Bobezeau
Vanessa
V
Hervouet
Nadège
N
Kellenberger
Christine
C
Roussel
Alain
A
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D003460:Crystallization; D018360:Crystallography, X-Ray; D029721:Drosophila Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D004331:Drosophila melanogaster / Q000737:chemistry*; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary
D002352:Carrier Proteins; D029721:Drosophila Proteins; C516487:GNBP3 protein, Drosophila; D047908:Intracellular Signaling Peptides and Proteins