Determination of adenosine deaminase binding domain on CD26 and its immunoregulatory effect on T cell activation.
CD26, a 110-kDa cell surface glycoprotein, exhibits dipeptidyl peptidase IV enzyme activity and plays an important role in T cell costimulation. In the present study, we examined both the exact adenosine deaminase (ADA) binding domain on CD26 and the functional consequences of mutated CD26 transfectants that were deficient for cell surface ADA. Using CD26 deletion, human-rat swap, and point mutations, we found that the residues of L340, V341, A342, and R343 on the CD26 molecule were essential amino acids for ADA binding. When these amino acids were mutated and transfected into Jurkat cells, the resultant CD26 transfectants expressed only CD26, not ADA, on the cell surface. The amount of IL-2 produced by wild-type and mutated CD26 transfectants was almost the same following stimulation with anti-CD3 plus PMA. However, the mutated CD26 transfectants were much more sensitive to the inhibitory effect of adenosine on IL-2 production than were the wild CD26 transfectants. These data suggest that ADA on the cell surface does not directly involve T cell activation. Conversely, CD26 alone does not result in modulating the inhibitory effect of adenosine. Only the ADA bound to CD26 on the cell surface was functional and could counteract the inhibitory effect of elevated extracellular adenosine.
Dong
R P
RP
Department of Medicine, Harvard Medical School, Boston, MA 02115, USA.
Tachibana
K
K
Hegen
M
M
Munakata
Y
Y
Cho
D
D
Schlossman
S F
SF
Morimoto
C
C
D000243:Adenosine Deaminase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D000276:Adjuvants, Immunologic / Q000378:metabolism / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000276:immunology; D018819:Dipeptidyl Peptidase 4 / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D017353:Gene Deletion; D006801:Humans; D019169:Jurkat Cells; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D051381:Rats; D013601:T-Lymphocytes / Q000201:enzymology* / Q000276:immunology*; D014162:Transfection / Q000276:immunology
D000276:Adjuvants, Immunologic; D018819:Dipeptidyl Peptidase 4; D000243:Adenosine Deaminase