Independent and synergistic effects of disulfide bond formation, beta 2-microglobulin, and peptides on class I MHC folding and assembly in an in vitro translation system.
We have examined the post-translational processing, intrachain disulfide bond formation, folding, and assembly of MHC class I H chains with beta 2-microglobulin after coupled in vitro translation of homogeneous mRNA and transport of nascent chains into canine microsomal vesicles. The formation of native alpha 3 domain conformation was dependent on conditions that optimized intrachain disulfide bond formation, and efficient folding of the alpha 1 alpha 2 domain required exposure to antigenic peptide. beta 2-microglobulin and peptide acted synergistically in forming native alpha 1 alpha 2 domain structure, and a small proportion of molecules with native alpha 1 alpha 2, but non-native alpha 3 structure were detected, indicating that alpha 3 domain folding is not an absolute prerequisite for the formation of native alpha 1 alpha 2 domain structure.
Ribaudo
R K
RK
Molecular Biology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
Margulies
D H
DH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004357:Drug Synergism; D004591:Electrophoresis, Polyacrylamide Gel; D015683:Gene Products, gag / Q000494:pharmacology*; D005978:Glutathione / Q000494:pharmacology; D006183:H-2 Antigens / Q000096:biosynthesis*; D064532:Histocompatibility Antigen H-2D; D017874:Immediate-Early Proteins; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008861:Microsomes / Q000502:physiology; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis / Q000187:drug effects; D011499:Protein Processing, Post-Translational; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000494:pharmacology*; D001613:beta 2-Microglobulin / Q000494:pharmacology*; D054301:gag Gene Products, Human Immunodeficiency Virus
D015683:Gene Products, gag; D006183:H-2 Antigens; D064532:Histocompatibility Antigen H-2D; D017874:Immediate-Early Proteins; D014764:Viral Proteins; D001613:beta 2-Microglobulin; C052612:cytomegalovirus immediate early phosphoprotein pp89; D054301:gag Gene Products, Human Immunodeficiency Virus; C061169:p18 gag protein, Human immunodeficiency virus 1; D005978:Glutathione