Crystallization and preliminary crystallographic analysis of the cell-surface alginate-binding protein Algp7 isolated from Sphingomonas sp. A1.

Sphingomonas sp. A1, a Gram-negative bacterium, directly internalizes alginate macromolecules through a mouth-like pit that is present on its cell surface. The alginate-binding protein Algp7, which was found to be localized on the cell surface, contributes to the accumulation of alginate in the pit. Algp7 was crystallized at 293 K by means of the sitting-drop vapour-diffusion method with polyethylene glycol 3350 as a crystallizing agent. Preliminary X-ray analysis showed that the Algp7 crystal belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 50.1, b = 98.0, c = 100.1 A, and that it diffracted to 2.8 A resolution.

[1]  K. Murata,et al.  A putative lipoprotein of Sphingomonas sp. strain A1 binds alginate rather than a lipid moiety. , 2008, FEMS microbiology letters.

[2]  B. Mikami,et al.  Superchannel of Bacteria: Biological Significance and New Horizons , 2008, Bioscience, biotechnology, and biochemistry.

[3]  B. Mikami,et al.  Proteomics-based identification of outer-membrane proteins responsible for import of macromolecules in Sphingomonas sp. A1: alginate-binding flagellin on the cell surface. , 2005, Biochemistry.

[4]  Wataru Hashimoto,et al.  Direct evidence for Sphingomonas sp. A1 periplasmic proteins as macromolecule-binding proteins associated with the ABC transporter: molecular insights into alginate transport in the periplasm. , 2005, Biochemistry.

[5]  Wataru Hashimoto,et al.  Molecular identification and characterization of an alginate-binding protein on the cell surface of Sphingomonas sp. A1. , 2004, Biochemical and biophysical research communications.

[6]  S. Kawai,et al.  Molecular Identification of Oligoalginate Lyase ofSphingomonas sp. Strain A1 as One of the Enzymes Required for Complete Depolymerization of Alginate , 2000, Journal of bacteriology.

[7]  K. Murata,et al.  A Novel Bacterial ATP-Binding Cassette Transporter System That Allows Uptake of Macromolecules , 2000, Journal of bacteriology.

[8]  B. Mikami,et al.  Overexpression in Escherichia coli, purification, and characterization of Sphingomonas sp. A1 alginate lyases. , 2000, Protein expression and purification.

[9]  Z. Otwinowski,et al.  [20] Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[10]  K. Murata,et al.  Pit structure on bacterial cell surface. , 1996, Biochemical and biophysical research communications.

[11]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[12]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[13]  B. Matthews Solvent content of protein crystals. , 1968, Journal of molecular biology.