Purification strategies for sendai virus membrane proteins

✍ Scribed by Gjalt W. Welling; Kunja Slopsema; Sytske Welling-Wester

• Book ID: 104145828
• Publisher: Elsevier Science
• Year: 1987
• Tongue: English
• Weight: 834 KB
• Volume: 397
• Category: Article
• ISSN: 1873-3778
• DOI: 10.1016/s0021-9673(01)84999-2

✦ Synopsis

Viral membrane proteins extracted from Sendai virions with the non-ionic detergents decylpolyethyleneglycol-300 and Triton X-100 were used as a model mixture of hydrophobic membrane proteins. The detergent extract contained the fusion protein (F) and the tetrameric and dimeric forms of the hemagglutinin-neuraminidase protein (HN). These proteins were purified by size-exclusion high-performance liquid chromatography (HPLC) in the presence of 0.1% sodium dodecyl sulphate, by ion-exchange and metal chelate affinity HPLC in the presence of 0.1% decylpolyethyleneglycol, and by reversed-phase HPLC without prior removal of the detergent. The tetramer of HN and F could be purified by size-exclusion HPLC after dissociation of a micellar aggregate containing tetrameric HN and multimeric F. The F and HN proteins could be purified by ion-exchange HPLC. Pure F protein could be obtained after metal chelate affinity HPLC. The F protein and the dimer and tetramer of HN could be eluted from a large-pore (100 nm) reversed-phase column, but they were eluted as broad, overlapping peaks. Only after reduction of the virion extract, the relatively small (13-15 kilodaltons) F2 protein could be obtained in pure form.