Abstract
A longstanding goal in the fields of molecular genetics and biochemistry has been to explain how naturally occurring mutations associated with human metabolic disease impair activity of the enzymes involved. This goal is particularly complex for enzymes composed of multiple subunits, because single mutations may exert both intra- and intersubunit effects on holoenzyme structure and function. We have previously applied a yeast coexpression system for human galactose-1-phosphate uridylyltransferase, a dimeric enzyme associated with galactosemia, to investigate the impact of naturally occurring mutations on subunit association and holoenzyme function (). Here we describe the purification and characterization of two heterodimers, R333W/wild type (WT) and Q188R/WT, revealing that although the first exhibits approximately 50% wild-type activity, the second exhibits only approximately 15% wild-type activity. Neither heterodimer varied significantly from the wild type with regard to apparent Km for either substrate, although Q188R/WT but not R333W/WT heterodimers demonstrated significantly increased thermal sensitivity relative to the wild-type enzyme. These results demonstrate for the first time a partial dominant negative effect caus...Continue Reading
References
Jan 1, 1978·Annual Review of Biochemistry·P Y Chou, G D Fasman
Nov 1, 1978·Archives of Biochemistry and Biophysics·V P Williams
Nov 27, 1990·Biochemistry·J KimP A Frey
Jan 1, 1989·Journal of Inherited Metabolic Disease·M BrivetA Lemonnier
Jan 1, 1989·Journal of Inherited Metabolic Disease·E PloechlK M Gibson
Feb 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·R GentzC A Rosen
Jan 1, 1989·The British Journal of Ophthalmology·R E StevensW J Stark
Oct 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·H L NadlerM Rachmeler
Feb 13, 1981·Biochimica Et Biophysica Acta·J BanroquesF Schapira
Sep 1, 1981·Archives of Biochemistry and Biophysics·G R Helmer, V P Williams
Jan 1, 1982·Methods in Enzymology·P A FreyS L Yang
Jan 1, 1983·Biochimie·J BanroquesJ C Dreyfus
Feb 1, 1980·Archives of Ophthalmology·H W Skalka, J T Prchal
Sep 5, 1995·Biochemistry·J E WedekindI Rayment
Apr 25, 1995·Biochemistry·F J RuzickaP A Frey
Mar 1, 1994·Human Genetics·A BoschX Estivill
May 18, 1994·Biochimica Et Biophysica Acta·L LinT S Ma
Jan 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·J L Fridovich-Keil, S Jinks-Robertson
Jul 9, 1996·Proceedings of the National Academy of Sciences of the United States of America·J P ElsevierJ L Fridovich-Keil
Dec 1, 1995·Biochemical and Molecular Medicine·J L Fridovich-KeilJ P Elsevier
Oct 25, 1996·The Journal of Biological Chemistry·B B QuimbyJ L Fridovich-Keil
Citations
Jul 17, 1999·Human Mutation·L TyfieldM D Boleda
Jan 23, 2008·Journal of Inherited Metabolic Disease·J S ChhayJ L Fridovich-Keil
Oct 22, 2008·Journal of Inherited Metabolic Disease·Johannes Zschocke
Jan 16, 2013·Journal of Inherited Metabolic Disease·Emily L RyanJudith L Fridovich-Keil
Sep 5, 2002·Molecular Genetics and Metabolism·Nicole C Christacos, Judith L Fridovich-Keil
Dec 17, 2009·Protein Engineering, Design & Selection : PEDS·A Facchiano, A Marabotti
Jul 11, 2006·Plant Physiology·Nir DaiArthur A Schaffer
Jun 4, 2010·Disease Models & Mechanisms·Rebekah F KushnerJudith L Fridovich-Keil
Oct 1, 2014·Archives of Physiology and Biochemistry·Peter SchadewaldtUdo Wendel
Oct 28, 2009·IUBMB Life·Kent LaiKlaas J Wierenga
Oct 1, 2011·IUBMB Life·Thomas J McCorvie, David J Timson
Apr 16, 2013·Biochimica Et Biophysica Acta·Thomas J McCorvieDavid J Timson
Jul 6, 2015·Gene·David J Timson
Feb 15, 2011·Biochimica Et Biophysica Acta·João LeandroTorgeir Flatmark
Feb 26, 1999·The Journal of Biological Chemistry·K LaiL J Elsas
Jan 21, 2001·The Journal of Biological Chemistry·K RiehmanJ L Fridovich-Keil
Nov 5, 1997·Journal of Inherited Metabolic Disease·L Wells, J L Fridovich-Keil
Dec 7, 2019·Journal of Inherited Metabolic Disease·Minela HaskovicM Estela Rubio-Gozalbo
Oct 30, 2016·Human Molecular Genetics·Thomas J McCorvieWyatt W Yue
Feb 4, 2005·Journal of Medicinal Chemistry·Anna Marabotti, Angelo M Facchiano
Oct 17, 1998·Biochemistry·S Geeganage, P A Frey
Aug 12, 1998·Biochemistry·F J RuzickaP A Frey
Mar 23, 2001·Genetics in Medicine : Official Journal of the American College of Medical Genetics·L J Elsas, K Lai