Purification and Composition of the Proteins from Sindbis Virus Grown in Chick and BHK Cells

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 1976 Dec 1

PMID 994303

Citations 20

Authors

D J Burke

K Keegstra

Affiliations

Soon will be listed here.

Abstract

Procedures are described for the purification of the Sindbis virus structural proteins. The amino acid and carbohydrate compositions of the purified proteins are presented for virus grown in BHK-21/13 and chicken embryo cells. Glycoprotein E1 from virus grown in BHK cells is deficient in a mannose-rich glycopeptide found on that glycoprotein when virus is grown in chicken embryo cells. The complex glactose-containing glycopeptides appear similar for virus grown in both hosts. However, when virus is grown in BHK cells, both glycoproteins are enriched in those glycopeptides containing more sialic acid. Since the two viral glycoproteins are difficult to separate cleanly during purification, it is suggested that there may be strong, but noncovalent, interactions between glycoproteins E1 and E2. It is also suggested that there may be an interaction between glycoprotein E2 and a component of the nucleocapsid.

Citing Articles

Encapsidation of host-derived factors correlates with enhanced infectivity of Sindbis virus.

Sokoloski K, Snyder A, Liu N, Hayes C, Mukhopadhyay S, Hardy R J Virol. 2013; 87(22):12216-26.

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Single amino acid insertions at the junction of the sindbis virus E2 transmembrane domain and endodomain disrupt virus envelopment and alter infectivity.

Hernandez R, Ferreira D, Sinodis C, Litton K, Brown D J Virol. 2005; 79(12):7682-97.

PMID: 15919921 PMC: 1143637. DOI: 10.1128/JVI.79.12.7682-7697.2005.

Sindbis virus glycoprotein E1 is divided into two discrete domains at amino acid 129 by disulfide bridge connections.

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PMID: 10982379 PMC: 102131. DOI: 10.1128/jvi.74.19.9313-9316.2000.

Purification of the envelope glycoproteins of western equine encephalitis virus by glass wool column chromatography.

Yamamoto K, Simizu B Appl Environ Microbiol. 1980; 40(2):240-3.

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One extra oligosaccharide chain of the high-mannose class in the E2 protein of a Sindbis virus isolate.

Cancedda R, Bonatti S, Leone A J Virol. 1981; 38(1):8-14.

PMID: 7241662 PMC: 171119. DOI: 10.1128/JVI.38.1.8-14.1981.

References

Elgin S, Weintraub H . Chromosomal proteins and chromatin structure. Annu Rev Biochem. 1975; 44:725-74. DOI: 10.1146/annurev.bi.44.070175.003453. View

Wang C, Smith R . Lowry determination of protein in the presence of Triton X-100. Anal Biochem. 1975; 63(2):414-7. DOI: 10.1016/0003-2697(75)90363-2. View

Keegstra K, Sefton B, Burke D . Sindbis virus glycoproteins: effect of the host cell on the oligosaccharides. J Virol. 1975; 16(3):613-20. PMC: 354709. DOI: 10.1128/JVI.16.3.613-620.1975. View

Gahmberg C, Utermann G, Simons K . The membrane proteins of Semliki Forest virus have a hydrophobic part attached to the viral membrane. FEBS Lett. 1972; 28(2):179-182. DOI: 10.1016/0014-5793(72)80706-3. View

Warren L . The thiobarbituric acid assay of sialic acids. J Biol Chem. 1959; 234(8):1971-5. View

Spiro R . THE CARBOHYDRATE UNITS OF THYROGLOBULIN. J Biol Chem. 1965; 240:1603-10. View

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

Brown D, Smith J . Morphology of BHK-21 Cells Infected with Sindbis Virus Temperature-Sensitive Mutants in Complementation Groups D and E. J Virol. 1975; 15(5):1262-6. PMC: 354582. DOI: 10.1128/JVI.15.5.1262-1266.1975. View

Schlesinger S, Gottlieb C, Feil P, Gelb N, Kornfeld S . Growth of enveloped RNA viruses in a line of chinese hamster ovary cells with deficient N-acetylglucosaminyltransferase activity. J Virol. 1975; 17(1):239-46. PMC: 515408. DOI: 10.1128/JVI.17.1.239-246.1976. View

10.

Helenius A, Simons K . The binding of detergents to lipophilic and hydrophilic proteins. J Biol Chem. 1972; 247(11):3656-61. View

11.

DAVID A . Lipid composition of Sindbis virus. Virology. 1971; 46(3):711-20. DOI: 10.1016/0042-6822(71)90073-0. View

12.

Etchison J, Holland J . Carbohydrate composition of the membrane glycoprotein of vesicular stomatitis virus. Virology. 1974; 60(1):217-29. DOI: 10.1016/0042-6822(74)90379-1. View

13.

Etchison J, Holland J . Carbohydrate composition of the membrane glycoprotein of vesicular stomatitis virus grown in four mammalian cell lines. Proc Natl Acad Sci U S A. 1974; 71(10):4011-4. PMC: 434317. DOI: 10.1073/pnas.71.10.4011. View

14.

MUDD J . Glycoprotein fragment associated with vesicular stomatitis virus after proteolytic digestion. Virology. 1974; 62(2):573-7. DOI: 10.1016/0042-6822(74)90419-x. View

15.

Garoff H, Simons K, Renkonen O . Isolation and characterization of the membrane proteins of Semliki Forest virus. Virology. 1974; 61(2):493-504. DOI: 10.1016/0042-6822(74)90285-2. View

16.

Hirschberg C, Robbins P . The glycolipids and phospholipids of Sindbis virus and their relation to the lipids of the host cell plasma membrane. Virology. 1974; 61(2):602-8. DOI: 10.1016/0042-6822(74)90295-5. View

17.

Rej R, Richards A . Interference by Tris buffer in the estimation of protein by the Lowry procedure. Anal Biochem. 1974; 62(1):240-7. DOI: 10.1016/0003-2697(74)90383-2. View

18.

Utermann G, Simons K . Studies on the amphipathic nature of the membrane proteins in Semliki Forest virus. J Mol Biol. 1974; 85(4):569-87. DOI: 10.1016/0022-2836(74)90316-7. View

19.

Brown D, Waite M, Pfefferkorn E . Morphology and morphogenesis of Sindbis virus as seen with freeze-etching techniques. J Virol. 1972; 10(3):524-36. PMC: 356494. DOI: 10.1128/JVI.10.3.524-536.1972. View

20.

Simmons D, Strauss J . Replication of Sindbis virus. I. Relative size and genetic content of 26 s and 49 s RNA. J Mol Biol. 1972; 71(3):599-613. View