NMR structure of phospho-tyrosine signaling complexes.
A structural basis for activation and substrate specificity of src tyrosine kinases, and regulation of protein-protein association by tyrosine phosphorylation is described. Lyn, a src-family tyrosine kinase, recognizes and phosphorylates the immunoreceptor tyrosine-based activation motif, ITAM, a critical component in transmembrane signal transduction in hemopoietic cells. The structure of an ITAM peptide substrate bound to an active form of Lyn tyrosine kinase was determined by high-resolution NMR, and a model of the complex was generated using the crystallographic structure of Lck, a closely related Src-family kinase. The results provide a rationale for the conserved ITAM residues and specificity of Lyn, and suggest that substrate plays a role in stabilizing the kinase conformation optimal for catalysis. It is our hope that the Lck-ITAM peptide model complex will be useful in aiding structure-based drug design efforts that target substrate binding determinants in the design. Concerning the regulation of protein-protein association, we report on a complex between erythrocyte band 3 and two glycolytic enzymes, aldolase and glyceraldehyde-3-phosphate dehydrogenase. The formation of this complex is negatively regulated by tyrosine phosphorylation of band 3 by p72syk tyrosine kinase. In red blood cells, this association results in a decrease in glycolysis due to competitive inhibition of the glycolytic enzymes. The structure of band 3 recognized by the glycolytic enzymes was determined by solution NMR, and found to be a loop structure with tyrosine centrally positioned and excluded from intermolecular contact. This phosphorylation sensitive interaction, or PSI, loop may be the basis of a general mechanism for negative regulation through tyrosine phosphorylation.
Post
C B
CB
Department of Medicinal Chemistry, Purdue University, West Lafayette IN 47907-1333, USA. cpb@stella.bio.purdue.edu
Gaul
B S
BS
Eisenmesser
E Z
EZ
Schneider
M L
ML
D015195:Drug Design; D006801:Humans; D008961:Models, Structural; D019000:Phosphotyrosine / Q000737:chemistry* / Q000502:physiology*; D011487:Protein Conformation; D021382:Protein Sorting Signals / Q000737:chemistry* / Q000502:physiology*
D021382:Protein Sorting Signals; D019000:Phosphotyrosine