Molecular dissection of the CD2-CD58 counter-receptor interface identifies CD2 Tyr86 and CD58 Lys34 residues as the functional "hot spot".
The heterophilic CD2-CD58 adhesion interface contains interdigitating residues that impart high specificity and rapid binding kinetics. To define the hot spot of this counter-receptor interaction, we characterized CD2 adhesion domain variants harboring a single mutation of the central Tyr86 or of each amino acid residue forming a salt link/hydrogen bond. Alanine mutations at D31, D32 and K34 on the C strand and K43 and R48 on the C' strand reduce affinity for CD58 by 47-127-fold as measured by isothermal titration calorimetry. The Y86A mutant reduces affinity by approximately 1000-fold, whereas Y86F is virtually without effect, underscoring the importance of the phenyl ring rather than the hydroxyl moiety. The CD2-CD58 crystal structure offers a detailed view of this key functional epitope: CD2 D31 and D32 orient the side-chain of CD58 K34 such that CD2 Y86 makes hydrophobic contact with the extended aliphatic component of CD58 K34 between CD2 Y86 and CD58 F46. The elucidation of this hot spot provides a new target for rational design of immunosuppressive compounds and suggests a general approach for other receptors.
Kim
M
M
Laboratory of Immunobiology, Dana-Farber Cancer Institute, 44 Binney Street, Boston, MA 02115, USA.
Sun
Z Y
ZY
Byron
O
O
Campbell
G
G
Wagner
G
G
Wang
J
J
Reinherz
E L
EL
D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D018801:CD2 Antigens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018968:CD58 Antigens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002151:Calorimetry; D002448:Cell Adhesion; D002850:Chromatography, Gel; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D006860:Hydrogen Bonding; D020128:Inhibitory Concentration 50; D019169:Jurkat Cells; D008239:Lysine / Q000235:genetics / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012397:Rosette Formation; D012756:Sheep; D055672:Static Electricity; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism*
D018801:CD2 Antigens; D018968:CD58 Antigens; D014443:Tyrosine; D008239:Lysine