title: Structure-function analysis of the bacterial expansin EXLX1.
abstract: We made use of EXLX1, an expansin from Bacillus subtilis, to investigate protein features essential for its plant cell wall binding and wall loosening activities. We found that the two expansin domains, D1 and D2, need to be linked for wall extension activity and that D2 mediates EXLX1 binding to whole cell walls and to cellulose via distinct residues on the D2 surface. Binding to cellulose is mediated by three aromatic residues arranged linearly on the putative binding surface that spans D1 and D2. Mutation of these three residues to alanine eliminated cellulose binding and concomitantly eliminated wall loosening activity measured either by cell wall extension or by weakening of filter paper but hardly affected binding to whole cell walls, which is mediated by basic residues located on other D2 surfaces. Mutation of these basic residues to glutamine reduced cell wall binding but not wall loosening activities. We propose domain D2 as the founding member of a new carbohydrate binding module family, CBM63, but its function in expansin activity apparently goes beyond simply anchoring D1 to the wall. Several polar residues on the putative binding surface of domain D1 are also important for activity, most notably Asp82, whose mutation to alanine or asparagine completely eliminated wall loosening activity. The functional insights based on this bacterial expansin may be extrapolated to the interactions of plant expansins with cell walls.
authors: Department of Biology, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors: Nikolaos
authors: N
authors: Georgelis
authors: Akira
authors: A
authors: Tabuchi
authors: Nikolas
authors: N
authors: Nikolaidis
authors: Daniel J
authors: DJ
authors: Cosgrove
mesh_terms: D000409:Alanine / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry* / Q000502:physiology; D001665:Binding Sites; D002473:Cell Wall / Q000378:metabolism; D002482:Cellulose / Q000737:chemistry; D003001:Cloning, Molecular; D041821:Gluconacetobacter / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010940:Plant Proteins / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship
chemical_list: D001426:Bacterial Proteins; D010940:Plant Proteins; D011134:Polysaccharides; C095825:expansin protein, plant; D002482:Cellulose; D000409:Alanine