B

Bálint Bécsi

University of Debrecen

Publishes on Protein Kinase Regulation and GTPase Signaling, Blood properties and coagulation, Muscle Physiology and Disorders. 32 papers and 541 citations.

32Publications
541Total Citations
Protein Kinase Regulation and GTPase SignalingBlood properties and coagulationMuscle Physiology and DisordersErythrocyte Function and PathophysiologyCellular Mechanics and Interactions

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Transglutaminase 2 Is Needed for the Formation of an Efficient Phagocyte Portal in Macrophages Engulfing Apoptotic Cells
Beáta B. Tóth, Éva Garabuczi, Zsolt Sarang et al.|The Journal of Immunology|2009
Cited by 140Open Access

Transglutaminase 2 (TG2), a protein cross-linking enzyme with many additional biological functions, acts as coreceptor for integrin beta(3). We have previously shown that TG2(-/-) mice develop an age-dependent autoimmunity due to defective in vivo clearance of apoptotic cells. Here we report that TG2 on the cell surface and in guanine nucleotide-bound form promotes phagocytosis. Besides being a binding partner for integrin beta(3), a receptor known to mediate the uptake of apoptotic cells via activating Rac1, we also show that TG2 binds MFG-E8 (milk fat globulin EGF factor 8), a protein known to bridge integrin beta(3) to apoptotic cells. Finally, we report that in wild-type macrophages one or two engulfing portals are formed during phagocytosis of apoptotic cells that are characterized by accumulation of integrin beta(3) and Rac1. In the absence of TG2, integrin beta(3) cannot properly recognize the apoptotic cells, is not accumulated in the phagocytic cup, and its signaling is impaired. As a result, the formation of the engulfing portals, as well as the portals formed, is much less efficient. We propose that TG2 has a novel function to stabilize efficient phagocytic portals.

Affinity, Avidity, and Kinetics of Target Sequence Binding to LC8 Dynein Light Chain Isoforms
László Radnai, Péter Rapali, Zsuzsa Hódi et al.|Journal of Biological Chemistry|2010
Cited by 37Open Access

LC8 dynein light chain (DYNLL) is a highly conserved eukaryotic hub protein with dozens of binding partners and various functions beyond being a subunit of dynein and myosin Va motor proteins. Here, we compared the kinetic and thermodynamic parameters of binding of both mammalian isoforms, DYNLL1 and DYNLL2, to two putative consensus binding motifs (KXTQTX and XG(I/V)QVD) and report only subtle differences. Peptides containing either of the above motifs bind to DYNLL2 with micromolar affinity, whereas a myosin Va peptide (lacking the conserved Gln) and the noncanonical Pak1 peptide bind with Kd values of 9 and 40 μm, respectively. Binding of the KXTQTX motif is enthalpy-driven, although that of all other peptides is both enthalpy- and entropy-driven. Moreover, the KXTQTX motif shows strikingly slower off-rate constant than the other motifs. As most DYNLL partners are homodimeric, we also assessed the binding of bivalent ligands to DYNLL2. Compared with monovalent ligands, a significant avidity effect was found as follows: Kd values of 37 and 3.5 nm for a dimeric myosin Va fragment and a Leu zipper dimerized KXTQTX motif, respectively. Ligand binding kinetics of DYNLL can best be described by a conformational selection model consisting of a slow isomerization and a rapid binding step. We also studied the binding of the phosphomimetic S88E mutant of DYNLL2 to the dimeric myosin Va fragment, and we found a significantly lower apparent Kd value (3 μm). We conclude that the thermodynamic and kinetic fine-tuning of binding of various ligands to DYNLL could have physiological relevance in its interaction network. LC8 dynein light chain (DYNLL) is a highly conserved eukaryotic hub protein with dozens of binding partners and various functions beyond being a subunit of dynein and myosin Va motor proteins. Here, we compared the kinetic and thermodynamic parameters of binding of both mammalian isoforms, DYNLL1 and DYNLL2, to two putative consensus binding motifs (KXTQTX and XG(I/V)QVD) and report only subtle differences. Peptides containing either of the above motifs bind to DYNLL2 with micromolar affinity, whereas a myosin Va peptide (lacking the conserved Gln) and the noncanonical Pak1 peptide bind with Kd values of 9 and 40 μm, respectively. Binding of the KXTQTX motif is enthalpy-driven, although that of all other peptides is both enthalpy- and entropy-driven. Moreover, the KXTQTX motif shows strikingly slower off-rate constant than the other motifs. As most DYNLL partners are homodimeric, we also assessed the binding of bivalent ligands to DYNLL2. Compared with monovalent ligands, a significant avidity effect was found as follows: Kd values of 37 and 3.5 nm for a dimeric myosin Va fragment and a Leu zipper dimerized KXTQTX motif, respectively. Ligand binding kinetics of DYNLL can best be described by a conformational selection model consisting of a slow isomerization and a rapid binding step. We also studied the binding of the phosphomimetic S88E mutant of DYNLL2 to the dimeric myosin Va fragment, and we found a significantly lower apparent Kd value (3 μm). We conclude that the thermodynamic and kinetic fine-tuning of binding of various ligands to DYNLL could have physiological relevance in its interaction network. IntroductionLC8 dynein light chain (DYNLL) 3The abbreviations used are: DYNLLLC8 dynein light chainCSconformational selectionDICdynein intermediate chainIFinduced fitITCisothermal titration calorimetrymyoVamyosin VaSPRsurface plasmon resonancenNOSnitric-oxide synthaseBmfBcl-2-modifying factorBimBcl-2 interacting mediator. is a highly conserved small eukaryotic protein. It was originally discovered as a light chain of the dynein (1King S.M. Patel-King R.S. J. Biol. Chem. 1995; 270: 11445-11452Abstract Full Text Full Text PDF PubMed Scopus (166) Google Scholar) and later of myosin Va (myoVa) (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar) motor protein complexes. However, DYNLL has many interaction partners unrelated to motor proteins. Therefore, it has been suggested that DYNLL is a hub protein that plays important roles in the interactome of eukaryotic cells in various cellular events, including apoptosis, molecular, organelle, and nuclear transport, viral infection, cancer development, and transcription regulation (3Barbar E. Biochemistry. 2008; 47: 503-508Crossref PubMed Scopus (134) Google Scholar, 4Hodi Z. Rapali P. Radnai L. Molnar T. Szenes A. Kardos J. Buday L. Stafford W. Nyitray L. FEBS J. 2007; 274 (106): 106Google Scholar). More intensively studied DYNLL-binding proteins include neuronal nitric-oxide synthase (nNOS) (5Jaffrey S.R. Snyder S.H. Science. 1996; 274: 774-777Crossref PubMed Scopus (423) Google Scholar), myoVa (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar), Bcl-2-modifying factor (Bmf) (6Puthalakath H. Villunger A. O'Reilly L.A. Beaumont J.G. Coultas L. Cheney R.E. Huang D.C. Strasser A. Science. 2001; 293: 1829-1832Crossref PubMed Scopus (497) Google Scholar), Bcl-2 interacting mediator (Bim) (7Puthalakath H. Huang D.C. O'Reilly L.A. King S.M. Strasser A. Mol. Cell. 1999; 3: 287-296Abstract Full Text Full Text PDF PubMed Scopus (904) Google Scholar), dynein intermediate chain (DIC) (8Makokha M. Hare M. Li M. Hays T. Barbar E. Biochemistry. 2002; 41: 4302-4311Crossref PubMed Scopus (88) Google Scholar), the Drosophila swallow mRNA localizing protein (9Schnorrer F. Bohmann K. Nüsslein-Volhard C. Nat. Cell Biol. 2000; 2: 185-190Crossref PubMed Scopus (207) Google Scholar), and p21-activated protein kinase 1 (Pak1) (10Vadlamudi R.K. Bagheri-Yarmand R. Yang Z. Balasenthil S. Nguyen D. Sahin A.A. den Hollander P. Kumar R. Cancer Cell. 2004; 5: 575-585Abstract Full Text Full Text PDF PubMed Scopus (158) Google Scholar, 11Lu J. Sun Q. Chen X. Wang H. Hu Y. Gu J. Biochem. Biophys. Res. Commun. 2005; 331: 153-158Crossref PubMed Scopus (12) Google Scholar). Several solution and crystal structures of apo-DYNLL and complexes with binding peptides have been determined (12Tochio H. Ohki S. Zhang Q. Li M. Zhang M. Nat. Struct. Biol. 1998; 5: 965-969Crossref PubMed Scopus (45) Google Scholar, 13Liang J. Jaffrey S.R. Guo W. Snyder S.H. Clardy J. Nat. Struct. Biol. 1999; 6: 735-740Crossref PubMed Scopus (143) Google Scholar, 14Fan J. Zhang Q. Tochio H. Li M. Zhang M. J. Mol. Biol. 2001; 306: 97-108Crossref PubMed Scopus (122) Google Scholar, 15Williams J.C. Roulhac P.L. Roy A.G. Vallee R.B. Fitzgerald M.C. Hendrickson W.A. Proc. Natl. Acad. Sci. U.S.A. 2007; 104: 10028-10033Crossref PubMed Scopus (111) Google Scholar, 16Benison G. Karplus P.A. Barbar E. J. Mol. Biol. 2007; 371: 457-468Crossref PubMed Scopus (70) Google Scholar, 17Benison G. Karplus P.A. Barbar E. J. Mol. Biol. 2008; 384: 954-966Crossref PubMed Scopus (36) Google Scholar, 18Wang W. Lo K.W. Kan H.M. Fan J.S. Zhang M. J. Biol. Chem. 2003; 278: 41491-41499Abstract Full Text Full Text PDF PubMed Scopus (39) Google Scholar, 19Day C.L. Puthalakath H. Skea G. Strasser A. Barsukov I. Lian L.Y. Huang D.C. Hinds M.G. Biochem. J. 2004; 377: 597-605Crossref PubMed Google Scholar, 20Makokha M. Huang Y.J. Montelione G. Edison A.S. Barbar E. Protein Sci. 2004; 13: 727-734Crossref PubMed Scopus (34) Google Scholar). DYNLL has a homodimeric structure, and the bound partner peptides lie in two identical grooves formed at the dimerization interface (12Tochio H. Ohki S. Zhang Q. Li M. Zhang M. Nat. Struct. Biol. 1998; 5: 965-969Crossref PubMed Scopus (45) Google Scholar, 13Liang J. Jaffrey S.R. Guo W. Snyder S.H. Clardy J. Nat. Struct. Biol. 1999; 6: 735-740Crossref PubMed Scopus (143) Google Scholar, 14Fan J. Zhang Q. Tochio H. Li M. Zhang M. J. Mol. Biol. 2001; 306: 97-108Crossref PubMed Scopus (122) Google Scholar, 15Williams J.C. Roulhac P.L. Roy A.G. Vallee R.B. Fitzgerald M.C. Hendrickson W.A. Proc. Natl. Acad. Sci. U.S.A. 2007; 104: 10028-10033Crossref PubMed Scopus (111) Google Scholar, 16Benison G. Karplus P.A. Barbar E. J. Mol. Biol. 2007; 371: 457-468Crossref PubMed Scopus (70) Google Scholar, 17Benison G. Karplus P.A. Barbar E. J. Mol. Biol. 2008; 384: 954-966Crossref PubMed Scopus (36) Google Scholar, 18Wang W. Lo K.W. Kan H.M. Fan J.S. Zhang M. J. Biol. Chem. 2003; 278: 41491-41499Abstract Full Text Full Text PDF PubMed Scopus (39) Google Scholar, 19Day C.L. Puthalakath H. Skea G. Strasser A. Barsukov I. Lian L.Y. Huang D.C. Hinds M.G. Biochem. J. 2004; 377: 597-605Crossref PubMed Google Scholar, 20Makokha M. Huang Y.J. Montelione G. Edison A.S. Barbar E. Protein Sci. 2004; 13: 727-734Crossref PubMed Scopus (34) Google Scholar). Formerly, it was widely assumed that DYNLL could function as a cargo adapter on dynein and myoVa motors (6Puthalakath H. Villunger A. O'Reilly L.A. Beaumont J.G. Coultas L. Cheney R.E. Huang D.C. Strasser A. Science. 2001; 293: 1829-1832Crossref PubMed Scopus (497) Google Scholar, 21Naisbitt S. Valtschanoff J. Allison D.W. Sala C. Kim E. Craig A.M. Weinberg R.J. Sheng M. J. Neurosci. 2000; 20: 4524-4534Crossref PubMed Google Scholar, 22Pfister K.K. Fisher E.M. Gibbons I.R. Hays T.S. Holzbaur E.L. McIntosh J.R. Porter M.E. Schroer T.A. Vaughan K.T. Witman G.B. King S.M. Vallee R.B. J. Cell Biol. 2005; 171: 411-413Crossref PubMed Scopus (157) Google Scholar). However, this hypothesis is difficult to reconcile with the symmetric homodimeric structure of DYNLL and most of its partners, including myoVa and DIC. Instead, it has been suggested, based on the effect of DYNLL on its partner proteins, that one of the major roles of DYNLL dimers could be the ability to promote dimerization and stabilization of their interaction partners (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar, 3Barbar E. Biochemistry. 2008; 47: 503-508Crossref PubMed Scopus (134) Google Scholar, 23Wang L. Hare M. Hays T.S. Barbar E. Biochemistry. 2004; 43: 4611-4620Crossref PubMed Scopus (58) Google Scholar).DYNLL has two mammalian isoforms (DYNLL1 and DYNLL2; previously known as DLC1 and DLC2 or LC8a and LC8b) (21Naisbitt S. Valtschanoff J. Allison D.W. Sala C. Kim E. Craig A.M. Weinberg R.J. Sheng M. J. Neurosci. 2000; 20: 4524-4534Crossref PubMed Google Scholar, 22Pfister K.K. Fisher E.M. Gibbons I.R. Hays T.S. Holzbaur E.L. McIntosh J.R. Porter M.E. Schroer T.A. Vaughan K.T. Witman G.B. King S.M. Vallee R.B. J. Cell Biol. 2005; 171: 411-413Crossref PubMed Scopus (157) Google Scholar) that differ from each other only in six residues. All of these residues are located outside of the ligand binding grooves. Despite their similarity, DYNLL1 and DYNLL2 seem to discriminate binding partners in the cell (6Puthalakath H. Villunger A. O'Reilly L.A. Beaumont J.G. Coultas L. Cheney R.E. Huang D.C. Strasser A. Science. 2001; 293: 1829-1832Crossref PubMed Scopus (497) Google Scholar, 19Day C.L. Puthalakath H. Skea G. Strasser A. Barsukov I. Lian L.Y. Huang D.C. Hinds M.G. Biochem. J. 2004; 377: 597-605Crossref PubMed Google Scholar), although some in vitro studies do not support this finding (21Naisbitt S. Valtschanoff J. Allison D.W. Sala C. Kim E. Craig A.M. Weinberg R.J. Sheng M. J. Neurosci. 2000; 20: 4524-4534Crossref PubMed Google Scholar, 24Lo K.W. Kogoy J.M. Rasoul B.A. King S.M. Pfister K.K. J. Biol. Chem. 2007; 282: 36871-36878Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar). The binding grooves are able to interact with short linear sequences, all of which are part of intrinsically of the partner proteins. binding motifs the based on KXTQTX and noncanonical myoVa and (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar, 14Fan J. Zhang Q. Tochio H. Li M. Zhang M. J. Mol. Biol. 2001; 306: 97-108Crossref PubMed Scopus (122) Google Scholar, I. F. F. FEBS 2001; PubMed Scopus Google Scholar, Sun S. T. J.C. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar, K.W. S. Fan J.S. Sheng M. Zhang M. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus (158) Google Scholar). However, the relevance of this has been of of various binding peptides and protein values nm and determined by (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar, Sun S. T. J.C. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar, J. A. Barbar E. Biochemistry. 2008; 47: PubMed Scopus Google Scholar, C. W. Chen M. J. Zhang M. Kumar R. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus (45) Google Scholar, W. E. A. Biochemistry. 2006; 45: PubMed Scopus Google Scholar, A. Hare M. Hays T.S. Barbar E. Biochemistry. 2004; 43: PubMed Scopus Google Scholar). However, it is important to that many of the partners to as and DYNLL most complexes with its partners (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar, 23Wang L. Hare M. Hays T.S. Barbar E. Biochemistry. 2004; 43: 4611-4620Crossref PubMed Scopus (58) Google Scholar). the of peptides not be used to the interaction with dimeric partners, which in are bivalent protein ligands J.C. Roulhac P.L. Roy A.G. Vallee R.B. Fitzgerald M.C. Hendrickson W.A. Proc. Natl. Acad. Sci. U.S.A. 2007; 104: 10028-10033Crossref PubMed Scopus (111) Google Scholar). we have previously of dimeric myoVa binding to DYNLL2, compared with a peptide (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar), and the was for a dimeric fragment Sun S. T. J.C. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar). However, the the and the of the binding partner was not in of the of DYNLL as a hub protein is not Binding of the partners to DYNLL could be by of or residues the DYNLL-binding motif K. R.J. Proc. Natl. Acad. Sci. U.S.A. 2003; PubMed Scopus Google Scholar). of of DYNLL could be of regulation by the to the the binding grooves C. W. Chen M. J. Zhang M. Kumar R. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus (45) Google Scholar, Y. G. A. Hays T.S. Barbar E. J. Biol. Chem. 2007; 282: Full Text Full Text PDF PubMed Scopus Google Scholar). It is not which kinase is in this Pak1 was originally to DYNLL (10Vadlamudi R.K. Bagheri-Yarmand R. Yang Z. Balasenthil S. Nguyen D. Sahin A.A. den Hollander P. Kumar R. Cancer Cell. 2004; 5: 575-585Abstract Full Text Full Text PDF PubMed Scopus (158) Google Scholar, Z. R.K. Kumar R. J. Biol. Chem. 2005; Full Text Full Text PDF PubMed Scopus Google a not support its Sun S. T. J.C. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus Google we report the kinetic and thermodynamic parameters of binding of DYNLL isoforms to various partners with monovalent and bivalent motifs. We found that the is by the bivalent of the binding We also found that the binding of both monovalent and bivalent ligands can be best by a conformational selection we that ligands can bind to the S88E phosphomimetic of DYNLL by the to the IntroductionLC8 dynein light chain (DYNLL) 3The abbreviations used are: DYNLLLC8 dynein light chainCSconformational selectionDICdynein intermediate chainIFinduced fitITCisothermal titration calorimetrymyoVamyosin VaSPRsurface plasmon resonancenNOSnitric-oxide synthaseBmfBcl-2-modifying factorBimBcl-2 interacting mediator. is a highly conserved small eukaryotic protein. It was originally discovered as a light chain of the dynein (1King S.M. Patel-King R.S. J. Biol. Chem. 1995; 270: 11445-11452Abstract Full Text Full Text PDF PubMed Scopus (166) Google Scholar) and later of myosin Va (myoVa) (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar) motor protein complexes. However, DYNLL has many interaction partners unrelated to motor proteins. Therefore, it has been suggested that DYNLL is a hub protein that plays important roles in the interactome of eukaryotic cells in various cellular events, including apoptosis, molecular, organelle, and nuclear transport, viral infection, cancer development, and transcription regulation (3Barbar E. Biochemistry. 2008; 47: 503-508Crossref PubMed Scopus (134) Google Scholar, 4Hodi Z. Rapali P. Radnai L. Molnar T. Szenes A. Kardos J. Buday L. Stafford W. Nyitray L. FEBS J. 2007; 274 (106): 106Google Scholar). More intensively studied DYNLL-binding proteins include neuronal nitric-oxide synthase (nNOS) (5Jaffrey S.R. Snyder S.H. Science. 1996; 274: 774-777Crossref PubMed Scopus (423) Google Scholar), myoVa (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar), Bcl-2-modifying factor (Bmf) (6Puthalakath H. Villunger A. O'Reilly L.A. Beaumont J.G. Coultas L. Cheney R.E. Huang D.C. Strasser A. Science. 2001; 293: 1829-1832Crossref PubMed Scopus (497) Google Scholar), Bcl-2 interacting mediator (Bim) (7Puthalakath H. Huang D.C. O'Reilly L.A. King S.M. Strasser A. Mol. Cell. 1999; 3: 287-296Abstract Full Text Full Text PDF PubMed Scopus (904) Google Scholar), dynein intermediate chain (DIC) (8Makokha M. Hare M. Li M. Hays T. Barbar E. Biochemistry. 2002; 41: 4302-4311Crossref PubMed Scopus (88) Google Scholar), the Drosophila swallow mRNA localizing protein (9Schnorrer F. Bohmann K. Nüsslein-Volhard C. Nat. Cell Biol. 2000; 2: 185-190Crossref PubMed Scopus (207) Google Scholar), and p21-activated protein kinase 1 (Pak1) (10Vadlamudi R.K. Bagheri-Yarmand R. Yang Z. Balasenthil S. Nguyen D. Sahin A.A. den Hollander P. Kumar R. Cancer Cell. 2004; 5: 575-585Abstract Full Text Full Text PDF PubMed Scopus (158) Google Scholar, 11Lu J. Sun Q. Chen X. Wang H. Hu Y. Gu J. Biochem. Biophys. Res. Commun. 2005; 331: 153-158Crossref PubMed Scopus (12) Google Scholar). Several solution and crystal structures of apo-DYNLL and complexes with binding peptides have been determined (12Tochio H. Ohki S. Zhang Q. Li M. Zhang M. Nat. Struct. Biol. 1998; 5: 965-969Crossref PubMed Scopus (45) Google Scholar, 13Liang J. Jaffrey S.R. Guo W. Snyder S.H. Clardy J. Nat. Struct. Biol. 1999; 6: 735-740Crossref PubMed Scopus (143) Google Scholar, 14Fan J. Zhang Q. Tochio H. Li M. Zhang M. J. Mol. Biol. 2001; 306: 97-108Crossref PubMed Scopus (122) Google Scholar, 15Williams J.C. Roulhac P.L. Roy A.G. Vallee R.B. Fitzgerald M.C. Hendrickson W.A. Proc. Natl. Acad. Sci. U.S.A. 2007; 104: 10028-10033Crossref PubMed Scopus (111) Google Scholar, 16Benison G. Karplus P.A. Barbar E. J. Mol. Biol. 2007; 371: 457-468Crossref PubMed Scopus (70) Google Scholar, 17Benison G. Karplus P.A. Barbar E. J. Mol. Biol. 2008; 384: 954-966Crossref PubMed Scopus (36) Google Scholar, 18Wang W. Lo K.W. Kan H.M. Fan J.S. Zhang M. J. Biol. Chem. 2003; 278: 41491-41499Abstract Full Text Full Text PDF PubMed Scopus (39) Google Scholar, 19Day C.L. Puthalakath H. Skea G. Strasser A. Barsukov I. Lian L.Y. Huang D.C. Hinds M.G. Biochem. J. 2004; 377: 597-605Crossref PubMed Google Scholar, 20Makokha M. Huang Y.J. Montelione G. Edison A.S. Barbar E. Protein Sci. 2004; 13: 727-734Crossref PubMed Scopus (34) Google Scholar). DYNLL has a homodimeric structure, and the bound partner peptides lie in two identical grooves formed at the dimerization interface (12Tochio H. Ohki S. Zhang Q. Li M. Zhang M. Nat. Struct. Biol. 1998; 5: 965-969Crossref PubMed Scopus (45) Google Scholar, 13Liang J. Jaffrey S.R. Guo W. Snyder S.H. Clardy J. Nat. Struct. Biol. 1999; 6: 735-740Crossref PubMed Scopus (143) Google Scholar, 14Fan J. Zhang Q. Tochio H. Li M. Zhang M. J. Mol. Biol. 2001; 306: 97-108Crossref PubMed Scopus (122) Google Scholar, 15Williams J.C. Roulhac P.L. Roy A.G. Vallee R.B. Fitzgerald M.C. Hendrickson W.A. Proc. Natl. Acad. Sci. U.S.A. 2007; 104: 10028-10033Crossref PubMed Scopus (111) Google Scholar, 16Benison G. Karplus P.A. Barbar E. J. Mol. Biol. 2007; 371: 457-468Crossref PubMed Scopus (70) Google Scholar, 17Benison G. Karplus P.A. Barbar E. J. Mol. Biol. 2008; 384: 954-966Crossref PubMed Scopus (36) Google Scholar, 18Wang W. Lo K.W. Kan H.M. Fan J.S. Zhang M. J. Biol. Chem. 2003; 278: 41491-41499Abstract Full Text Full Text PDF PubMed Scopus (39) Google Scholar, 19Day C.L. Puthalakath H. Skea G. Strasser A. Barsukov I. Lian L.Y. Huang D.C. Hinds M.G. Biochem. J. 2004; 377: 597-605Crossref PubMed Google Scholar, 20Makokha M. Huang Y.J. Montelione G. Edison A.S. Barbar E. Protein Sci. 2004; 13: 727-734Crossref PubMed Scopus (34) Google Scholar). Formerly, it was widely assumed that DYNLL could function as a cargo adapter on dynein and myoVa motors (6Puthalakath H. Villunger A. O'Reilly L.A. Beaumont J.G. Coultas L. Cheney R.E. Huang D.C. Strasser A. Science. 2001; 293: 1829-1832Crossref PubMed Scopus (497) Google Scholar, 21Naisbitt S. Valtschanoff J. Allison D.W. Sala C. Kim E. Craig A.M. Weinberg R.J. Sheng M. J. Neurosci. 2000; 20: 4524-4534Crossref PubMed Google Scholar, 22Pfister K.K. Fisher E.M. Gibbons I.R. Hays T.S. Holzbaur E.L. McIntosh J.R. Porter M.E. Schroer T.A. Vaughan K.T. Witman G.B. King S.M. Vallee R.B. J. Cell Biol. 2005; 171: 411-413Crossref PubMed Scopus (157) Google Scholar). However, this hypothesis is difficult to reconcile with the symmetric homodimeric structure of DYNLL and most of its partners, including myoVa and DIC. Instead, it has been suggested, based on the effect of DYNLL on its partner proteins, that one of the major roles of DYNLL dimers could be the ability to promote dimerization and stabilization of their interaction partners (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar, 3Barbar E. Biochemistry. 2008; 47: 503-508Crossref PubMed Scopus (134) Google Scholar, 23Wang L. Hare M. Hays T.S. Barbar E. Biochemistry. 2004; 43: 4611-4620Crossref PubMed Scopus (58) Google Scholar).DYNLL has two mammalian isoforms (DYNLL1 and DYNLL2; previously known as DLC1 and DLC2 or LC8a and LC8b) (21Naisbitt S. Valtschanoff J. Allison D.W. Sala C. Kim E. Craig A.M. Weinberg R.J. Sheng M. J. Neurosci. 2000; 20: 4524-4534Crossref PubMed Google Scholar, 22Pfister K.K. Fisher E.M. Gibbons I.R. Hays T.S. Holzbaur E.L. McIntosh J.R. Porter M.E. Schroer T.A. Vaughan K.T. Witman G.B. King S.M. Vallee R.B. J. Cell Biol. 2005; 171: 411-413Crossref PubMed Scopus (157) Google Scholar) that differ from each other only in six residues. All of these residues are located outside of the ligand binding grooves. Despite their similarity, DYNLL1 and DYNLL2 seem to discriminate binding partners in the cell (6Puthalakath H. Villunger A. O'Reilly L.A. Beaumont J.G. Coultas L. Cheney R.E. Huang D.C. Strasser A. Science. 2001; 293: 1829-1832Crossref PubMed Scopus (497) Google Scholar, 19Day C.L. Puthalakath H. Skea G. Strasser A. Barsukov I. Lian L.Y. Huang D.C. Hinds M.G. Biochem. J. 2004; 377: 597-605Crossref PubMed Google Scholar), although some in vitro studies do not support this finding (21Naisbitt S. Valtschanoff J. Allison D.W. Sala C. Kim E. Craig A.M. Weinberg R.J. Sheng M. J. Neurosci. 2000; 20: 4524-4534Crossref PubMed Google Scholar, 24Lo K.W. Kogoy J.M. Rasoul B.A. King S.M. Pfister K.K. J. Biol. Chem. 2007; 282: 36871-36878Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar). The binding grooves are able to interact with short linear sequences, all of which are part of intrinsically of the partner proteins. binding motifs the based on KXTQTX and noncanonical myoVa and (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar, 14Fan J. Zhang Q. Tochio H. Li M. Zhang M. J. Mol. Biol. 2001; 306: 97-108Crossref PubMed Scopus (122) Google Scholar, I. F. F. FEBS 2001; PubMed Scopus Google Scholar, Sun S. T. J.C. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar, K.W. S. Fan J.S. Sheng M. Zhang M. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus (158) Google Scholar). However, the relevance of this has been of of various binding peptides and protein values nm and determined by (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar, Sun S. T. J.C. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar, J. A. Barbar E. Biochemistry. 2008; 47: PubMed Scopus Google Scholar, C. W. Chen M. J. Zhang M. Kumar R. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus (45) Google Scholar, W. E. A. Biochemistry. 2006; 45: PubMed Scopus Google Scholar, A. Hare M. Hays T.S. Barbar E. Biochemistry. 2004; 43: PubMed Scopus Google Scholar). However, it is important to that many of the partners to as and DYNLL most complexes with its partners (2Hódi Z. Németh A.L. Radnai L. Hetényi C. Schlett K. Bodor A. Perczel A. Nyitray L. Biochemistry. 2006; 45: 12582-12595Crossref PubMed Scopus (49) Google Scholar, 23Wang L. Hare M. Hays T.S. Barbar E. 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U.S.A. 2003; PubMed Scopus Google Scholar). of of DYNLL could be of regulation by the to the the binding grooves C. W. Chen M. J. Zhang M. Kumar R. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus (45) Google Scholar, Y. G. A. Hays T.S. Barbar E. J. Biol. Chem. 2007; 282: Full Text Full Text PDF PubMed Scopus Google Scholar). It is not which kinase is in this Pak1 was originally to DYNLL (10Vadlamudi R.K. Bagheri-Yarmand R. Yang Z. Balasenthil S. Nguyen D. Sahin A.A. den Hollander P. Kumar R. Cancer Cell. 2004; 5: 575-585Abstract Full Text Full Text PDF PubMed Scopus (158) Google Scholar, Z. R.K. Kumar R. J. Biol. Chem. 2005; Full Text Full Text PDF PubMed Scopus Google a not support its Sun S. T. J.C. J. Biol. Chem. 2008; Full Text Full Text PDF PubMed Scopus Google we report the kinetic and thermodynamic parameters of binding of DYNLL isoforms to various partners with monovalent and bivalent motifs. We found that the is by the bivalent of the binding We also found that the binding of both monovalent and bivalent ligands can be best by a conformational selection we that ligands can bind to the S88E phosphomimetic of DYNLL by the to the

Epigallocatechin‐3‐gallate and penta‐<i>O</i>‐galloyl‐β‐d‐glucose inhibit protein phosphatase‐1
Andrea Kiss, Bálint Bécsi, Bernadett Kolozsvári et al.|FEBS Journal|2012
Cited by 31Open Access

Protein phosphatase-1 (PP1) and protein phosphatase-2A (PP2A) are responsible for the dephosphorylation of the majority of phosphoserine/threonine residues in cells. In this study, we show that (-)-epigallocatechin-3-gallate (EGCG) and 1,2,3,4,6-penta-O-galloyl-β-D-glucose (PGG), polyphenolic constituents of green tea and tannins, inhibit the activity of the PP1 recombinant δ-isoform of the PP1 catalytic subunit and the native PP1 catalytic subunit (PP1c) with IC(50) values of 0.47-1.35 μm and 0.26-0.4 μm, respectively. EGCG and PGG inhibit PP2Ac less potently, with IC(50) values of 15 and 6.6 μm, respectively. The structure-inhibitory potency relationships of catechin derivatives suggests that the galloyl group may play a major role in phosphatase inhibition. The interaction of EGCG and PGG with PP1c was characterized by NMR and surface plasmon resonance-based binding techniques. Competitive binding assays and molecular modeling suggest that EGCG docks at the hydrophobic groove close to the catalytic center of PP1c, partially overlapping with the binding surface of microcystin-LR or okadaic acid. This hydrophobic interaction is further stabilized by hydrogen bonding via hydroxyl/oxo groups of EGCG to PP1c residues. Comparative docking shows that EGCG binds to PP2Ac in a similar manner, but in a distinct pose. Long-term treatment (24 h) with these compounds and other catechins suppresses the viability of HeLa cells with a relative effectiveness reminiscent of their in vitro PP1c-inhibitory potencies. The above data imply that the phosphatase-inhibitory features of these polyphenols may be implicated in the wide spectrum of their physiological influence.