Total chemical synthesis, characterization, and immunological properties of an MHC class I model using the TASP concept for protein de novo design

Details

Serval ID
serval:BIB_50CB74C58DCF
Type
Article: article from journal or magazin.
Collection
Publications
Title
Total chemical synthesis, characterization, and immunological properties of an MHC class I model using the TASP concept for protein de novo design
Journal
Protein Science
Author(s)
Tuchscherer  G., Servis  C., Corradin  G., Blum  U., Rivier  J., Mutter  M.
ISSN
0961-8368 (Print)
Publication state
Published
Issued date
10/1992
Volume
1
Number
10
Pages
1377-86
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Oct
Abstract
The design, total chemical synthesis, and immunological properties of a four-alpha-helix bundle template-assembled synthetic protein (TASP) mimicking some of the structural features of the major histocompatibility complex (MHC) class I is described. In a first approach, the native sequence 58-74 of the alpha 1 heavy chain domain of HLA-A2 was modeled in order to increase helix stability and amphiphilicity of the 17-mer peptide, preserving the residues for potential T-cell receptor (TcR) binding properties. According to the TASP concept, these helical segments were covalently attached to a cyclic template molecule designed for the induction of a four-helix-bundle topology of the assembled peptide blocks. After extensive HPLC purification, stepwise solid-phase synthesis resulted in a TASP molecule of high chemical purity as demonstrated by analytical HPLC, mass spectrometry, and amino acid analysis. CD spectroscopic investigations are consistent with the onset of a partial alpha-helical conformation in aqueous buffer as well as in TFE. Antibodies raised directly against this four-alpha-helix bundle TASP molecule (without prior conjugation to a carrier molecule) were detected by ELISA. Flow cytometry studies showed that these antibodies recognize the native MHC class I molecule on the surface of HLA-A2-positive cells. The results indicate that the TASP approach represents a versatile tool for mimicking conformational epitopes.
Keywords
Amino Acid Sequence Animals Antibodies/immunology Cell Line Circular Dichroism Female Flow Cytometry HLA-A2 Antigen/immunology Histocompatibility Antigens Class I/*chemistry/immunology Mass Spectrometry Mice Mice, Inbred BALB C Mice, Inbred C57BL Molecular Sequence Data Protein Conformation
Pubmed
Web of science
Open Access
Yes
Create date
24/01/2008 14:55
Last modification date
20/08/2019 14:06
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