title: A subdomain interaction at the base of the lever allosterically tunes the mechanochemical mechanism of myosin 5a.
abstract: The motor domain of myosin is the core element performing mechanochemical energy transduction. This domain contains the actin and ATP binding sites and the base of the force-transducing lever. Coordinated subdomain movements within the motor are essential in linking the ATPase chemical cycle to translocation along actin filaments. A dynamic subdomain interface located at the base of the lever was previously shown to exert an allosteric influence on ATP hydrolysis in the non-processive myosin 2 motor. By solution kinetic, spectroscopic and ensemble and single-molecule motility experiments, we determined the role of a class-specific adaptation of this interface in the mechanochemical mechanism of myosin 5a, a processive intracellular transporter. We found that the introduction of a myosin 2-specific repulsive interaction into myosin 5a via the I67K mutation perturbs the strong-binding interaction of myosin 5a with actin, influences the mechanism of ATP binding and facilitates ATP hydrolysis. At the same time, the mutation abolishes the actin-induced activation of ADP release and, in turn, slows down processive motility, especially when myosin experiences mechanical drag exerted by the action of multiple motor molecules bound to the same actin filament. The results highlight that subtle structural adaptations of the common structural scaffold of the myosin motor enable specific allosteric tuning of motor activity shaped by widely differing physiological demands.
authors: Department of Biochemistry, ELTE-MTA, Eötvös Loránd University-Hungarian Academy of Sciences, Momentum Motor Enzymology Research Group, Eötvös Loránd University, Budapest, Hungary.
authors: Nikolett T
authors: NT
authors: Nagy
authors: Saikat
authors: S
authors: Chakraborty
authors: Gábor M
authors: GM
authors: Harami
authors: James R
authors: JR
authors: Sellers
authors: Takeshi
authors: T
authors: Sakamoto
authors: Mihály
authors: M
authors: Kovács
mesh_terms: D000199:Actins / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D000494:Allosteric Regulation; D019943:Amino Acid Substitution; D000818:Animals; D001696:Biomechanical Phenomena; D006868:Hydrolysis; D007700:Kinetics; D051379:Mice; D018995:Myosin Heavy Chains / Q000737:chemistry* / Q000235:genetics; D024701:Myosin Type V / Q000737:chemistry* / Q000235:genetics; D010710:Phosphates / Q000737:chemistry; D017354:Point Mutation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs
chemical_list: D000199:Actins; C513082:Myo5a protein, mouse; D010710:Phosphates; D000255:Adenosine Triphosphate; D024701:Myosin Type V; D018995:Myosin Heavy Chains
references: Biophys J. 2004 Sep;87(3):1776-83
references: 15345556
references: Biochemistry. 1997 Sep 30;36(39):11828-36
references: 9305974
references: Nucleic Acids Res. 2010 Jul;38(13):4404-14
references: 20211839
references: Proc Natl Acad Sci U S A. 2010 Apr 13;107(15):6799-804
references: 20351242
references: FASEB J. 2010 Nov;24(11):4480-90
references: 20631329
references: Philos Trans R Soc Lond B Biol Sci. 2004 Dec 29;359(1452):1829-41
references: 15647159
references: Biochem J. 1974 Aug;141(2):351-64
references: 4281654
references: EMBO J. 2004 Nov 24;23(23):4527-37
references: 15510214
references: Trends Biochem Sci. 2010 Dec;35(12):684-90
references: 20801044
references: J Cell Biol. 1990 Aug;111(2):453-63
references: 2143195
references: Biochem J. 2000 Sep 15;350 Pt 3:693-9
references: 10970781
references: Biochemistry. 2002 Jul 2;41(26):8508-17
references: 12081502
references: J Biol Chem. 1971 Aug 10;246(15):4866-71
references: 4254541
references: Biophys J. 2012 Jun 6;102(11):2545-55
references: 22713570
references: Curr Opin Cell Biol. 2004 Feb;16(1):61-7
references: 15037306
references: FASEB J. 2011 Jan;25(1):111-21
references: 20837775
references: Biochemistry. 2005 Dec 13;44(49):16203-10
references: 16331980
references: Biochemistry. 2000 Dec 26;39(51):16135-46
references: 11123942
references: Biochemistry. 1994 Jul 12;33(27):8262-71
references: 8031761
references: Structure. 2007 May;15(5):553-64
references: 17502101
references: Adv Protein Chem. 2005;71:161-93
references: 16230112
references: Proc Natl Acad Sci U S A. 2000 Oct 10;97(21):11238-43
references: 11016966
references: Proc Natl Acad Sci U S A. 1999 Nov 23;96(24):13726-31
references: 10570140
references: Eur J Biochem. 1982 Nov 15;128(2-3):547-55
references: 7151795
references: Biochem J. 1988 Nov 15;256(1):41-6
references: 3223909
references: J Biol Chem. 2001 Jun 1;276(22):19483-90
references: 11278775
references: Biochemistry. 1995 Jul 18;34(28):8960-72
references: 7619795
references: Cytoskeleton (Hoboken). 2010 May;67(5):273-85
references: 20191566
references: Nat Struct Mol Biol. 2012 Feb 12;19(3):299-306
references: 22343723
references: Nature. 2003 Sep 25;425(6956):419-23
references: 14508494
references: J Biol Chem. 1991 Jan 5;266(1):294-302
references: 1845966
references: J Mol Biol. 2011 Apr 15;407(5):716-30
references: 21315083
references: Biochemistry. 2001 Oct 23;40(42):12727-37
references: 11601998
references: J Muscle Res Cell Motil. 1983 Apr;4(2):253-62
references: 6863518
references: Science. 2003 Jun 27;300(5628):2061-5
references: 12791999
references: J Biol Chem. 2007 Jun 15;282(24):17658-64
references: 17449872
references: J Biol Chem. 2003 Aug 1;278(31):29201-7
references: 12740393
references: Anal Biochem. 2009 Apr 1;387(1):20-9
references: 19154726