Biotecnologia Aplicada Elfos Scientiae ISSN: 0684-4551 Vol. 12, Num. 3, 1995, pp. 176

REPORTE CORTO / SHORT REPORT

X-RAY INVESTIGATION OF A RECOMBINANT OUTER MEMBRANE PROTEIN FROM Neisseria meningitidis

Ines Li de la Sierra^1,2, Thierry Prange^2,3, Pedro Saludjian^3, Marck Schiltz^2, Roger Fourme^2, Lucile Pernot^2 and Gabriel Padron^1.

^1CIGB, Ave. 31 entre 158 y 190, Cubanacan, La Habana, Cuba ^2LURE (CNRS, CEA, MESR), Batiment 209, Universite Paris-Sud, 91405-ORSAY CEDEX, France. ^3Chimie Biomoleculaire (URA 1430 du CNRS), UFR-Biomedicale, 93012-BOBIGNY CEDEX, France.

Code Number: BA95069
Sizes of Files:
    Text: 2K
    No associated graphics

This protein contains 599 amino acid residues, including six cysteine residues and a flavin adenosine dinucleotide (FAD) prosthetic group. The first 111 amino acid residues of the N-terminal domain are homologous with the lipoyl domain of acetyltranferases. The other segment of the sequence, which represents almost the total protein, displays high homology with lipoamide dehydrogenases, the flavoenzymes containing a redox-active disulfide.The p64K crystallizes in the space group P43212. Unit cell parameters are a=b=140 A, c=79 A. The structure of dihydrolipoamide domain of the P64K is solved by Molecular Replacement with the Navazas's package (AMORE). The know crystal structure of dihydrolipoamide dehydrogenase of Pseudomonas putida was used as the starting model.

The molecule was rotated and traslated in the asymmetric unit until obtention of the best rotation/translation parameters. The model was first refined by rigid body refinement at low resolution, then refined against 3 A data with the XPLOR program.The multiple isomorphous replacement method (MIR) is under evaluation to improve phases for location of the remaining 116 residues.

The optimization of the crystallization procedure (with NAD cofactor) will help to improve the diffraction and resolution.

Copyright 1995 Sociedad Iberolatinamericana de Biotecnologia Aplicada a la Salud