Slower processing, weaker beta 2-M association, and lower surface expression of H-2Ld are influenced by its amino terminus.
To determine why Ld antigens are expressed on the cell surface at levels three to four times lower than Dd or Kd antigens, pulse-chase experiments were used to compare their rates of biosynthesis and processing. Electrophoresis on sodium dodecyl sulfate gradient polyacrylamide gels resolved immunoprecipitates of each of these histocompatibility complex class I molecules into a slower and faster species. During the chase period, the faster migrating species appeared to be converted to the slower migrating species in a time-dependent manner. However, the conversion of Ld from the faster to the slower migrating species proceeded significantly more slowly than did the conversion of either Dd or Kd. Endoglycosidase H sensitivity and cell surface radiolabeling were used to determine the glycosylation state and cell location of each species of Ld and Dd. The results from these experiments, along with the pulse-chase studies and cytofluorometric analyses, suggest that Ld possesses a much slower rate of processing from a faster migrating, high mannose-bearing species to a slower migrating, complex oligosaccharide-bearing species found on the cell surface. Analysis of the beta 2-microglobulin (beta 2-m) association confirmed that Ld is associated with less beta 2-m than Dd. To localize the structures on class I molecules influencing their surface expression, rate of processing, and beta 2-m association, the Ddm1 molecule was analyzed. The Ddm1 molecule of the mutant B10.D2-H-2dm1 has previously been shown to be a chimeric Dd (amino-terminal)/Ld (carboxyl-terminal) polypeptide. The surface expression, processing and beta 2-m association of Ddm1 were found to be similar to Dd rather than Ld, suggesting that each of these phenomena are influenced by protein structure in the amino terminus.
D000595:Amino Acid Sequence; D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D006183:H-2 Antigens / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D064532:Histocompatibility Antigen H-2D; D051379:Mice; D008807:Mice, Inbred BALB C; D011499:Protein Processing, Post-Translational; D013329:Structure-Activity Relationship; D001613:beta 2-Microglobulin / Q000032:analysis*
D000954:Antigens, Surface; D006183:H-2 Antigens; D064532:Histocompatibility Antigen H-2D; D001613:beta 2-Microglobulin