Structural, energetic, and functional analysis of a protein-protein interface at distinct stages of affinity maturation.
Due to a paucity of studies that synthesize structural, energetic, and functional analyses of a series of protein complexes representing distinct stages in an affinity maturation pathway, the biophysical basis for the molecular evolution of protein-protein interactions is poorly understood. Here, we combine crystal structures and binding-free energies of a series of variant superantigen (SAG)-major histocompatibility complex (MHC) class II complexes exhibiting increasingly higher affinity to reveal that this affinity maturation pathway is controlled largely by two biophysical factors: shape complementarity and buried hydrophobic surface. These factors, however, do not contribute equivalently to the affinity maturation of the interface, as the former dominates the early steps of the maturation process while the latter is responsible for improved binding in later steps. Functional assays reveal how affinity maturation of the SAG-MHC interface corresponds to T cell activation by SAGs.
Sundberg
Eric J
EJ
Center for Advanced Research in Biotechnology, W.M. Keck Laboratory for Structural Biology, University of Maryland Biotechnology Institute, 9600 Gudelsky Drive, Rockville, MD 20850, USA. sundberg@umbi.umd.edu
Andersen
Peter S
PS
Schlievert
Patrick M
PM
Karjalainen
Klaus
K
Mariuzza
Roy A
RA
D001665:Binding Sites; D004768:Enterotoxins / Q000737:chemistry / Q000378:metabolism; D015798:HLA-DR1 Antigen / Q000737:chemistry / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000737:chemistry* / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D008213:Lymphocyte Activation; D008958:Models, Molecular; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D018089:Superantigens / Q000737:chemistry* / Q000378:metabolism*; D013601:T-Lymphocytes / Q000737:chemistry / Q000378:metabolism
D004768:Enterotoxins; D015798:HLA-DR1 Antigen; D000949:Histocompatibility Antigens Class II; D018089:Superantigens; C034959:enterotoxin C, staphylococcal