Versatile retargeting of SH3 domain binding by modification of non-conserved loop residues.
Src-homology (SH3) domain belongs to a class of ubiquitous modular protein domains found in nature. SH3 domains have a conserved surface that recognises proline-rich peptides in ligand proteins, but additional contacts also contribute to binding. Using the SH3 domain of hematopoietic cell kinase as a test case, we show that SH3 binding properties can be profoundly altered by modifications within a hexapeptide sequence in the RT-loop region that is not involved in recognition of currently known consensus SH3 target peptides. These results highlight the role of non-conserved regions in SH3 target selection, and introduce a strategy that may be generally feasible for generating artificial SH3 domains with desired ligand binding properties.
Hiipakka
Marita
M
Institute of Medical Technology, University of Tampere and Tampere University Hospital, Tampere, Finland.
Saksela
Kalle
K
D051722:ADAM Proteins / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites; D017252:CD3 Complex / Q000737:chemistry / Q000378:metabolism; D017124:Conserved Sequence; D015693:Gene Products, nef / Q000737:chemistry / Q000378:metabolism; D007118:Immunoassay / Q000379:methods; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D008954:Models, Biological; D008958:Models, Molecular; D017232:Oncogene Proteins v-mos / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism; D017433:Protein Structure, Secondary; D021381:Protein Transport; D020837:SOS1 Protein / Q000737:chemistry / Q000378:metabolism; D054462:p21-Activated Kinases; D018909:src Homology Domains
D017252:CD3 Complex; D015693:Gene Products, nef; D008565:Membrane Proteins; D017232:Oncogene Proteins v-mos; D020837:SOS1 Protein; C515903:PAK1 protein, human; D017346:Protein Serine-Threonine Kinases; D054462:p21-Activated Kinases; D051722:ADAM Proteins; C098660:ADAM15 protein, human