Disulfide bond formation of the in vitro-translated large antigen of hepatitis D virus.
The in vitro transcription and translation coupling system has been demonstrated to be a powerful method of characterizing protein encoded by a cloned gene. Two cDNA constructs coding hepatitis D viral (HDV) antigen, small (S) and large (L) DAg, respectively, were subjected to in vitro transcription and translation to examine multimer formation ability. By using 2-D-SDS-PAGE (non-reducing and reducing) analysis, two novel characteristics of the LDAg were found: (i) the forming of a homodimer and (ii) the formation of a complex with an unidentified 11-kDa protein via a disulfide bond. These features were neither found in SDAg nor in a cysteine-negative mutant of LDAg. Based on the fact of isoprenylation occurring at the sole cysteine residue of LDAg, it is suggested that the formation of a disulfide bond of LDAg might be involved in a transition toward isoprenylation.
Hu
H M
HM
Institute of Microbiology and Immunology, School of Life Sciences, National Yang-Ming University, Taipei, Taiwan, ROC.
Shih
K N
KN
Lo
S J
SJ
D003545:Cysteine; D004220:Disulfides; D018963:Hepatitis Antigens / Q000737:chemistry* / Q000235:genetics; D003698:Hepatitis Delta Virus / Q000737:chemistry* / Q000235:genetics; D035923:Hepatitis delta Antigens; D006801:Humans; D010084:Oxidation-Reduction; D014176:Protein Biosynthesis; D014764:Viral Proteins / Q000737:chemistry
D004220:Disulfides; D018963:Hepatitis Antigens; D035923:Hepatitis delta Antigens; D014764:Viral Proteins; C442244:hepatitis delta virus large antigen; D003545:Cysteine