Abstract:
:Protein engineering has been employed to investigate the effect of specific amino acid changes on the targeting of heterologous proteins to the outer cell surface of the Gram-positive bacterium Staphylococcus xylosus. Three different variants, corresponding to a 101 amino acid region of the major glycoprotein (G protein) of human respiratory syncytial virus (RSV), were generated in which multiple hydrophobic phenylalanine residues were either substituted or deleted. The different G protein fragments were expressed as one part of recombinant receptors designed for surface display on S. xylosus cells. The engineered variants of the RSV G protein hybrid receptors were, in contrast to a non-engineered fragment, efficiently targeted to the outer cell surface of recombinant S. xylosus cells as determined by different methods, including fluorescence-activated cell sorting. In addition, immunization of mice with live recombinant S. xylosus demonstrated that surface exposure was required to generate receptor-specific antibodies. The present strategy of hydrophobic engineering should be of general interest in surface-display applications and for secretion of proteins otherwise difficult to translocate through host cell membranes.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Nguyen TN,Gourdon MH,Hansson M,Robert A,Samuelson P,Libon C,Andréoni C,Nygren PA,Binz H,Uhlén Mdoi
10.1016/0168-1656(95)00081-zsubject
Has Abstract,Author List Incompletepub_date
1995-10-16 00:00:00pages
207-19issue
3eissn
0168-1656issn
1873-4863pii
016816569500081Zjournal_volume
42pub_type
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