The topology of the S protein in the yeast-derived hepatitis B surface antigen particles.
Hepatitis B surface antigen particles are highly immunogenic and have been shown to provide a suitable support for the presentation of foreign epitopes. More information about the topology of their constitutive protein, the S (small envelope) protein, is a prerequisite to any rational attempt to replace region of this protein with foreign epitopes without modifying the assembly of the particle. The topology of the S protein within the lipid membrane was investigated here by combining extensive proteolysis of the external protein domains with proteinase K and (FTIR-ATR). The proteolytic hydrolysis of the S protein and the identification of the digestion products allowed characterization of the membrane-protected regions of the protein. FTIR spectra of the digested hepatitis B particles revealed that the peptides associated with the particles are rich in alpha-helix structure. The kinetic of 2H/H exchange provided evidence that a large fraction of the native S protein is poorly accessible to the aqueous medium.
Sonveaux
N
N
Laboratoire de Chimie-Physique des Macromolécules aux Interfaces, Université Libre de Bruxelles, Belgium.
Conrath
K
K
Capiau
C
C
Brasseur
R
R
Goormaghtigh
E
E
Ruysschaert
J M
JM
D000626:Aminopeptidases / Q000378:metabolism; D003903:Deuterium; D019286:Endopeptidase K; D006514:Hepatitis B Surface Antigens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006868:Hydrolysis; D063208:Methionyl Aminopeptidases; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D017421:Sequence Analysis; D012697:Serine Endopeptidases / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared; D015003:Yeasts / Q000235:genetics
D006514:Hepatitis B Surface Antigens; D010446:Peptide Fragments; D011994:Recombinant Proteins; D003903:Deuterium; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases; D012697:Serine Endopeptidases; D019286:Endopeptidase K