Structure of the glycan chain from the surface layer glycoprotein of Clostridium thermohydrosulfuricum L77-66.
The thermophilic eubacterium Clostridium thermohydrosulfuricum L77-66 is covered by a crystalline surface layer composed of identical glycoprotein subunits which are arranged in a hexagonal lattice with centre-to-centre spacings of approx. 14.3 nm. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of cell wall preparations showed the presence of several broadened, carbohydrate-containing bands in a molecular mass range of 90 to 200 kDa. A total carbohydrate content of approx. 14% was determined in the purified surface layer glycoprotein. Chemical deglycosylation of this material by trifluoromethanesulfonic acid resulted in the disappearance of the complex banding pattern. Only a single band with a molecular mass of 82 kDa remained visible upon Coomassie staining. After proteolytic digestion of the surface layer glycoprotein a single glycopeptide fraction with an apparent molecular mass of approx. 25 kDa was obtained by gel filtration. Composition analysis, methylation, periodate oxidation and a combination of homonuclear and 1H-detected heteronuclear shift-correlated nuclear magnetic resonance experiments established the following structure for the glycan chain of the surface layer glycoprotein.
Altman
E
E
Institute for Biological Sciences, National Research Council, Ottawa, Ontario, Canada.
Brisson
J R
JR
Gagné
S M
SM
Kolbe
J
J
Messner
P
P
Sleytr
U B
UB
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D003013:Clostridium / Q000737:chemistry* / Q000648:ultrastructure; D005613:Freeze Etching; D008401:Gas Chromatography-Mass Spectrometry; D009682:Magnetic Resonance Spectroscopy; D008562:Membrane Glycoproteins / Q000737:chemistry*; D008698:Mesylates; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000302:isolation & purification*
D001425:Bacterial Outer Membrane Proteins; D008562:Membrane Glycoproteins; D008698:Mesylates; D011134:Polysaccharides; C012077:trifluoromethanesulfonic acid