High-resolution surface views of human lymphocytes during capping of CD4 and HLA antigens as revealed by immunogold fracture-flip.
The surface ultrastructure of lymphocytes during capping of two transmembrane proteins is shown. As seen by fracture-flip the plasma membranes of human lymphocytes are covered by a high density of surface particles. Incubation in 30% glycerol leads to aggregation of these surface particles. Immunogold labelling shows that the transmembrane proteins bearing HLA class I and CD4 antigens are confined to the particle aggregates. These results indicate that surface particles revealed by fracture-flip represent surface protrusions of integral membrane proteins seen as intramembrane particles in freeze-fractured lymphocytes. During capping HLA or CD4 antigens aggregate into progressively larger patches and, finally, into single caps. As revealed by fracture-flip the patches/caps are seen as clearly differentiated raised platforms that are clearly and sharply demarcated relative to contiguous areas of the surface. In non-patched (non-capped) regions, the pattern of distribution and apparent density of surface particles remain unaltered. Immunogold labelling clearly demarcates patches and caps, and shows that virtually no antigen molecules remain dispersed over the non-patched (non-capped) regions. Estimates of the surface density of either HLA or CD4 antigens over the capped areas point to high planar concentrations of the transmembrane proteins that bear these antigens.
Pavan
A
A
Dipartimento di Medicina Sperimentale, Università di Roma La Sapienza, Italy.
Mancini
P
P
Lucania
G
G
Frati
L
L
Torrisi
M R
MR
Pinto da Silva
P
P
D000954:Antigens, Surface / Q000378:metabolism*; D015704:CD4 Antigens / Q000378:metabolism*; D002462:Cell Membrane / Q000648:ultrastructure; D005614:Freeze Fracturing; D006680:HLA Antigens / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007152:Immunologic Capping / Q000502:physiology*; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000276:immunology / Q000648:ultrastructure*
D000954:Antigens, Surface; D015704:CD4 Antigens; D006680:HLA Antigens