title: A mechanism of global shape-dependent recognition and phosphorylation of filamin by protein kinase A.
abstract: Protein phosphorylation mediates essentially all aspects of cellular life. In humans, this is achieved by ∼500 kinases, each recognizing a specific consensus motif (CM) in the substrates. The majority of CMs are surface-exposed and are thought to be accessible to kinases for phosphorylation. Here we investigated the archetypical protein kinase A (PKA)-mediated phosphorylation of filamin, a major cytoskeletal protein that can adopt an autoinhibited conformation. Surprisingly, autoinhibited filamin is refractory to phosphorylation by PKA on a known Ser(2152) site despite its CM being exposed and the corresponding isolated peptide being readily phosphorylated. Structural analysis revealed that although the CM fits into the PKA active site its surrounding regions sterically clash with the kinase. However, upon ligand binding, filamin undergoes a conformational adjustment, allowing rapid phosphorylation on Ser(2152). These data uncover a novel ligand-induced conformational switch to trigger filamin phosphorylation. They further suggest a substrate shape-dependent filtering mechanism that channels specific exposed CM/kinase recognition in diverse signaling responses.
authors: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors: Sujay Subbayya
authors: SS
authors: Ithychanda
authors: Protein-Nucleic Acid Interaction Section, Structural Biophysics Laboratory, NCI, National Institutes of Health, Frederick, Maryland 21702, and.
authors: Xianyang
authors: X
authors: Fang
authors: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors: Maradumane L
authors: ML
authors: Mohan
authors: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195, Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio 44106.
authors: Liang
authors: L
authors: Zhu
authors: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors: Kalyan C
authors: KC
authors: Tirupula
authors: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors: Sathyamangla V
authors: SV
authors: Naga Prasad
authors: Protein-Nucleic Acid Interaction Section, Structural Biophysics Laboratory, NCI, National Institutes of Health, Frederick, Maryland 21702, and.
authors: Yun-Xing
authors: YX
authors: Wang
authors: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors: Sadashiva S
authors: SS
authors: Karnik
authors: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195, Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio 44106 qinj@ccf.org.
authors: Jun
authors: J
authors: Qin
mesh_terms: D000595:Amino Acid Sequence; D016384:Consensus Sequence; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry*; D064448:Filamins / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D010748:Phosphopeptides / Q000737:chemistry; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational
chemical_list: D064448:Filamins; D010748:Phosphopeptides; D017868:Cyclic AMP-Dependent Protein Kinases
keywords: Enzyme Catalysis; Filamin; Nuclear Magnetic Resonance (NMR); Protein Kinase A (PKA); Protein Phosphorylation; Serine/Threonine Protein Kinase