Expression, purification and preliminary crystallographic characterization of a novel segment from the neurofibromatosis type 1 protein.
Neurofibromin (MW 320 kDa) is the protein responsible for the pathogenesis of neurofibromatosis type 1 (NF1), one of the most common genetic diseases worldwide. The neurofibromin GAP-related domain (GRD, MW 38 kDa) possess a Ras-specific GTPase-activating protein property, which is at present its only clear biochemical function. This article describes the study of the bacterial production and preliminary X-ray crystallographic analysis of a neurofibromin fragment located at the C-terminal end of the GRD, which contains a region reported to be homologous to the yeast Sec14p lipid exchange protein. Of the three crystal variants obtained, a tetragonal form diffracted to a resolution of at least 2.3 A.
Bonneau
Fabien
F
European Molecular Biology Laboratory, Structural and Computational Biology Programme, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
D'Angelo
Igor
I
Welti
Stefan
S
Stier
Gunter
G
Ylänne
Jari
J
Scheffzek
Klaus
K
D000818:Animals; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004330:Drosophila / Q000378:metabolism; D017353:Gene Deletion; D006801:Humans; D008055:Lipids / Q000737:chemistry; D025542:Neurofibromin 1 / Q000737:chemistry*; D049294:Phospholipid Transfer Proteins / Q000737:chemistry; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry
D008055:Lipids; D025542:Neurofibromin 1; D049294:Phospholipid Transfer Proteins; C483438:SEC14 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins