Roles of structural domains in the morphology and surface anchoring of the tetragonal paracrystalline array of Aeromonas hydrophila. Biochemical characterization of the major structural domain.
The tetragonally arranged S-layer of Aeromonas hydrophila contains two morphological domains. The mature S-layer protein of A. hydrophila has a subunit molecular weight of 52,000, and has been reported to contain two structural domains. Here a mutant has been isolated which produces an S-layer of subunit molecular weight 38,650 as determined by sedimentation analysis. This truncated S-protein was exported via the periplasm to the cell surface, but could not self-assemble into a tetragonal array or be anchored to the cell surface. Instead the truncated protein formed cup-like structures which were purified and characterized biochemically. Automated Edman degradation showed that the truncated protein comprised the amino-terminal structural domain of the S-protein. This domain had an increased hydrophobic amino acid content relative to the wild-type protein, and contained approximately 42% beta-sheet, 10% alpha-helix, and 19% beta-turn. Differences in alpha-helix and beta-turn contents between the wild-type and truncated proteins were observed when the effects of pH and SDS were examined, indicating that the carboxy terminus influences the effects of environmental change on the conformation of the S-protein. This lesser carboxy-terminal array also appears to be required for both correct array morphology, and array anchoring, while the greater amino-terminal domain appears to comprise the major morphological core of the surface array.
Thomas
S
S
Department of Biochemistry and Microbiology, University of Victoria, B.C., Canada.
Austin
J W
JW
McCubbin
W D
WD
Kay
C M
CM
Trust
T J
TJ
D016980:Aeromonas hydrophila / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D000596:Amino Acids / Q000032:analysis; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D002458:Cell Fractionation; D002942:Circular Dichroism; D004251:DNA Transposable Elements; D008562:Membrane Glycoproteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D008854:Microscopy, Electron; D008970:Molecular Weight; D009154:Mutation; D017433:Protein Structure, Secondary
D000596:Amino Acids; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D008562:Membrane Glycoproteins; C489049:regular S-layer protein, bacteria