Sialic acid binding activity of transmissible gastroenteritis coronavirus affects sedimentation behavior of virions and solubilized glycoproteins.
The sedimentation behavior of transmissible gastroenteritis coronavirus (TGEV) was analyzed. Upon sucrose gradient centrifugation, the major virus band was found at a density of 1.20 to 1.22 g/cm(3). This high density was observed only when TGEV with a functional sialic acid binding activity was analyzed. Mutants of TGEV that lacked sialic acid binding activity due to a point mutation in the sialic acid binding site of the S protein were mainly recovered at a lower-density position on the sucrose gradient (1.18 to 1.19 g/cm(3)). Neuraminidase treatment of purified virions resulted in a shift of the sedimentation value from the higher to the lower density. These results suggest that binding of sialoglycoproteins to the virion surface is responsible for the sedimentation behavior of TGEV. When purified virions were treated with octylglucoside to solubilize viral glycoproteins, ultracentrifugation resulted in sedimentation of the S protein of TGEV. However, when neuraminidase-treated virions or mutants with a defective sialic acid binding activity were analyzed, the S protein remained in the supernatant rather than in the pellet fraction. These results indicate that the interaction of the surface protein S with sialoglycoconjugates is maintained after solubilization of this viral glycoprotein by detergent treatment.
Krempl
C
C
Institut für Virologie, Philipps-Universität Marburg, 35037 Marburg, Germany.
Herrler
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G
D000818:Animals; D002499:Centrifugation, Density Gradient / Q000379:methods; D005960:Glucosides / Q000494:pharmacology; D008854:Microscopy, Electron; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D009439:Neuraminidase / Q000378:metabolism; D012995:Solubility; D013552:Swine; D005760:Transmissible gastroenteritis virus / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism; D014771:Virion / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism
D005960:Glucosides; D014764:Viral Proteins; C064484:spike protein S, Transmissible gastroenteritis virus; C018619:octyl-beta-D-glucoside; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid