Rotational and lateral dynamics of I-A(k) molecules expressing cytoplasmic truncations.
Rotational and lateral diffusion of I-A(k) molecules with various alpha and beta chain cytoplasmic truncations known to affect class II function were measured to assess the role of cytoplasmic domains in regulating I-A(k) molecular motions. Deletion of all 12 alpha chain C-terminal residues and all 18 corresponding beta chain residues (alpha-12/beta-18) is known to abrogate translocation of protein kinase C to the nucleus upon class II cross-linking. Similarly, truncation of the entire cytoplasmic alpha chain domain and the 10 C-terminal residues of the beta chain impairs presentation of antigenic peptides to T cells. The rotational correlation time of the wild-type molecule, 11.9 +/- 2.6 micros as measured by time-resolved phosphorescence anisotropy, decreased to 7. 2 +/- 3.7 micros in the fully truncated alpha-12/beta-18 protein. Other truncated class II molecules exhibited only small changes in molecular rotation rates relative to the wild-type. The rate of lateral diffusion of the fully truncated molecule, measured with two independent methods, 2.3 x 10(-10) cm(2)/s, was comparable with that of the wild-type molecule. Thus, it appears that the alpha and beta chain cytoplasmic domains regulate the molecular motions of unperturbed I-A(k) molecules only modestly, despite the known involvement of these regions in class II signaling. Various explanations for this behavior are discussed, e.g. the possibility that class II membrane complexes are sufficiently large that association and dissociation of specific signaling proteins during antigen presentation do not significantly perturb the apparent molecular motions of the complex.
Munnelly
H M
HM
Department of Chemistry, Colorado State University, Fort Collins, CO 80523, USA.
Brady
C J
CJ
Hagen
G M
GM
Wade
W F
WF
Roess
D A
DA
Barisas
B G
BG
D016880:Anisotropy; D000911:Antibodies, Monoclonal / Q000276:immunology; D017951:Antigen Presentation; D002460:Cell Line; D000949:Histocompatibility Antigens Class II / Q000737:chemistry* / Q000276:immunology; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D008163:Luminescent Measurements; D008565:Membrane Proteins / Q000737:chemistry
D000911:Antibodies, Monoclonal; D000949:Histocompatibility Antigens Class II; D007140:Immunoglobulin Fab Fragments; D008565:Membrane Proteins