Cell fusion experiments reveal distinctly different association characteristics of cell-surface receptors.
The existence of small- and large-scale membrane protein clusters, containing dimers, oligomers and hundreds of proteins, respectively, has become widely accepted. However, it is largely unknown whether the internal structure of these formations is dynamic or static. Cell fusion was used to perturb the distribution of existing membrane protein clusters, and to investigate their mobility and associations. Scanning near-field optical microscopy, confocal and electron microscopy were applied to detect the exchange of proteins between large-scale protein clusters, whereas photobleaching fluorescence energy transfer was used to image the redistribution of existing small-scale membrane protein clusters. Large-scale clusters of major histocompatibility complex (MHC)-I exchanged proteins with each other and with MHC-II clusters. Similarly to MHC-I, large-scale MHC-II clusters were also dynamic. Exchange of components between small-scale protein clusters was not universal: intermixing did not take place in the case of MHC-II homoclusters; however, it was observed for homoclusters of MHC-I and for heteroclusters of MHC-I and MHC-II. These processes required a fluid state of the plasma membrane, and did not depend on endocytosis-mediated recycling of proteins. The redistribution of large-scale MHC-I clusters precedes the intermixing of small-scale clusters of MHC-I indicating a hierarchy in protein association. Investigation of a set of other proteins (alpha subunit of the interleukin 2 receptor, CD48 and transferrin receptor) suggested that a large-scale protein cluster usually exchanges components with the same type of clusters. These results offer new insight into processes requiring time-dependent changes in membrane protein interactions.
Nagy
P
P
Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary.
Mátyus
L
L
Jenei
A
A
Panyi
G
G
Varga
S
S
Matkó
J
J
Szöllosi
J
J
Gáspár
R
R
Jovin
T M
TM
Damjanovich
S
S
D002459:Cell Fusion; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D004735:Energy Transfer; D005456:Fluorescent Dyes / Q000378:metabolism; D017979:Gold Colloid / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D000949:Histocompatibility Antigens Class II / Q000378:metabolism*; D006801:Humans; D021962:Membrane Microdomains; D008853:Microscopy; D008856:Microscopy, Fluorescence / Q000379:methods; D011940:Receptor Aggregation / Q000502:physiology*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D015375:Receptors, Interleukin-2
D005456:Fluorescent Dyes; D017979:Gold Colloid; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D011956:Receptors, Cell Surface; D015375:Receptors, Interleukin-2