title: Structural and Dynamic Features of F-recruitment Site Driven Substrate Phosphorylation by ERK2.
abstract: The F-recruitment site (FRS) of active ERK2 binds F-site (Phe-x-Phe-Pro) sequences found downstream of the Ser/Thr phospho-acceptor on cellular substrates. Here we apply NMR methods to analyze the interaction between active ERK2 (ppERK2), and a 13-residue F-site-bearing peptide substrate derived from its cellular target, the transcription factor Elk-1. Our results provide detailed insight into previously elusive structural and dynamic features of FRS/F-site interactions and FRS-driven substrate phosphorylation. We show that substrate F-site engagement significantly quenches slow dynamics involving the ppERK2 activation-loop and the FRS. We also demonstrate that the F-site phenylalanines make critical contacts with ppERK2, in contrast to the proline whose cis-trans isomerization has no significant effect on F-site recognition by the kinase FRS. Our results support a mechanism where phosphorylation of the disordered N-terminal phospho-acceptor is facilitated by its increased productive encounters with the ppERK2 active site due to docking of the proximal F-site at the kinase FRS.
authors.affiliation: Department of Chemistry, The City College of New York, New York, NY 10031.
authors.forename: Andrea
authors.initials: A
authors.lastname: Piserchio
authors.affiliation: Department of Chemistry, The City College of New York, New York, NY 10031.
authors.forename: Venkatesh
authors.initials: V
authors.lastname: Ramakrishan
authors.affiliation: Department of Chemistry, The City College of New York, New York, NY 10031.
authors.forename: Hsin
authors.initials: H
authors.lastname: Wang
authors.affiliation: 1] Division of Medicinal Chemistry, University of Texas, Austin, TX 78712 [2] Department of Medicinal Chemistry, Faculty of Pharmacy, Minia University, El-Minia 61519, Egypt.
authors.forename: Tamer S
authors.initials: TS
authors.lastname: Kaoud
authors.affiliation: Department of Chemistry, The College of Staten Island, Staten Island, NY 10314.
authors.forename: Boris
authors.initials: B
authors.lastname: Arshava
authors.affiliation: The New York Structural Biology Center, New York, NY 10027.
authors.forename: Kaushik
authors.initials: K
authors.lastname: Dutta
authors.affiliation: Division of Medicinal Chemistry, University of Texas, Austin, TX 78712.
authors.forename: Kevin N
authors.initials: KN
authors.lastname: Dalby
authors.affiliation: 1] Department of Chemistry, The City College of New York, New York, NY 10031 [2] Graduate Center of CUNY, New York, NY 10016.
authors.forename: Ranajeet
authors.initials: R
authors.lastname: Ghose
mesh_terms: D001665:Binding Sites / Q000502:physiology; D020134:Catalytic Domain / Q000502:physiology*; D004247:DNA / Q000378:metabolism; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D008958:Models, Molecular; D010766:Phosphorylation; D011487:Protein Conformation; D050807:ets-Domain Protein Elk-1 / Q000378:metabolism*
chemical_list: C494112:ELK1 protein, human; D050807:ets-Domain Protein Elk-1; D004247:DNA; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1