A Dynamic molecular basis for malfunction in disease mutants of p97/VCP.
p97/VCP is an essential, abundant AAA+ ATPase that is conserved throughout eukaryotes, with central functions in diverse processes ranging from protein degradation to DNA damage repair and membrane fusion. p97 has been implicated in the etiology of degenerative diseases and in cancer. Using Nuclear Magnetic Resonance spectroscopy we reveal how disease-causing mutations in p97 deregulate dynamics of the N-terminal domain that binds adaptor proteins involved in controlling p97 function. Our results provide a molecular basis for understanding how malfunction occurs whereby mutations shift the ADP-bound form of the enzyme towards an ATP-like state in a manner that correlates with disease severity. This deregulation interferes with the two-pronged binding of an adaptor that affects p97 function in lysosomal degradation of substrates. Subtle structural changes propagate from mutation sites to regions distal in space, defining allosteric networks that facilitate inter-domain communication, with potential implications for modulation of enzyme activity by drug molecules.
Department of Molecular Genetics, University of Toronto, Toronto, Canada.|Department of Biochemistry, University of Toronto, Toronto, Cananda.|Department of Chemistry, University of Toronto, Toronto, Canada.
Anne K
AK
Schuetz
Department of Molecular Genetics, University of Toronto, Toronto, Canada.|Department of Biochemistry, University of Toronto, Toronto, Cananda.|Department of Chemistry, University of Toronto, Toronto, Canada.|Program in Molecular Structure and Function, Hospital for Sick Children, Toronto, Canada.
Lewis E
0000-0002-4054-4083
LE
Kay
D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D050505:Mutant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation
D050505:Mutant Proteins; D009687:Nuclear Proteins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C404942:p97 ATPase
IBMPFD disease mutations; biophysics; methyl-TROSY NMR; none; p97 adaptors; p97/VCP; protein dynamics, thermodynamics, equilibrium; structural biology