Abstract
Saccharomyces cerevisiae myristoyl-CoA:protein N-myristoyltransferase (Nmt1p) is an essential 455-residue, monomeric enzyme that catalyzes the transfer of myristate from myristoyl-CoA to the NH2-terminal Gly residue of cellular proteins. Nmt1p has an ordered Bi Bi reaction mechanism with binding of myristoyl-CoA occurring before binding of peptide substrates. To define residues important for function, the polymerase chain reaction was used to generate random mutations in the NMT1 gene. A colony color sectoring assay was used to screen a library of 52,000 transformants for nmt1 alleles encoding enzymes with reduced activity. nmt1 alleles were identified that produced temperature-sensitive (ts) growth arrest due to substitutions affecting eight residues conserved in orthologous Nmts: Asn102, Ala202, Cys217, Ser328, Val395, Asn404, Leu420, and Asn426. Ala202 --> Thr, Cys217 --> Arg, Ser328 --> Pro, Asn404 --> Tyr, and Asn426 --> Ile produced the most severe ts phenotype. Their effects on the functional properties of the enzyme's myristoyl-CoA and peptide binding sites were defined by purifying each mutant from Escherichia coli and conducting in vitro kinetic analyses with acyl-CoA and peptide substrates and with two competitive in...Continue Reading
References
Aug 1, 1992·Molecular and Cellular Biology·M S Cyert, J Thorner
Nov 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·D A RudnickJ I Gordon
Feb 1, 1992·Yeast·D MuhlradR Parker
May 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·R J DuronioJ I Gordon
May 1, 1992·Proteins·R J DuronioM S Boguski
Mar 1, 1992·The Journal of Cell Biology·D E Levin, E Bartlett-Heubusch
Nov 1, 1991·Genes & Development·D E StoneS I Reed
Mar 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·M L BryantJ I Gordon
Jun 1, 1991·The Journal of Cell Biology·R J DuronioJ I Gordon
Dec 1, 1990·Molecular and Cellular Biology·T StearnsM A Hoyt
Feb 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·R J DuronioJ I Gordon
Jul 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·R O Heuckeroth, J I Gordon
Feb 10, 1989·Science·R J DuronioJ I Gordon
May 1, 1989·Genetics·R S Sikorski, P Hieter
Aug 1, 1989·Journal of Medicinal Chemistry·L A PaigeR L Geahlen
Jan 1, 1987·Gene·C S Hoffman, F Winston
Feb 1, 1985·Cell·D KoshlandL H Hartwell
Feb 1, 1985·Cell·P HieterR W Davis
Feb 1, 1974·Journal of Bacteriology·K H Meyer, E Schweizer
Aug 15, 1970·Nature·U K Laemmli
Jan 1, 1983·Journal of Bacteriology·H ItoA Kimura
Oct 20, 1995·The Journal of Biological Chemistry·D ZhuJ Heitman
Nov 1, 1995·Journal of the National Cancer Institute·R L FelstedK Hartman
Nov 1, 1995·Journal of the National Cancer Institute·B A MagnusonR K Sharma
Apr 1, 1995·Molecular Microbiology·R A WeinbergS C Lee
Apr 1, 1995·Trends in Biochemical Sciences·A L Boman, R A Kahn
May 26, 1995·Journal of Medicinal Chemistry·B DevadasJ I Gordon
Jan 6, 1995·The Journal of Biological Chemistry·R A KahnM Cavenagh
Oct 11, 1994·Proceedings of the National Academy of Sciences of the United States of America·D R JohnsonJ I Gordon
Jan 1, 1994·Annual Review of Biochemistry·D R JohnsonJ I Gordon
Dec 6, 1994·Proceedings of the National Academy of Sciences of the United States of America·J K LodgeJ I Gordon
Sep 7, 1993·Biochemistry·M J NadlerR L Geahlen
Jun 1, 1993·The Journal of General Virology·D R HarperR A McIlhinney
Sep 1, 1993·Molecular and Cellular Biology·K S LeeD E Levin
Citations
Jan 1, 1997·Biopolymers·J A SikorskiJ I Gordon
Nov 10, 2009·Journal of Chemical Biology·Megan H WrightEdward W Tate
Dec 10, 1998·Nature Structural Biology·R S BhatnagarG Waksman
Mar 21, 1998·Nature Structural Biology·S A WestonR A Pauptit
Jun 5, 1998·Nucleic Acids Research·S J CokJ I Gordon
Nov 29, 2002·Eukaryotic Cell·Brian D ShawMichelle Momany
Aug 2, 2002·Expert Opinion on Investigational Drugs·Nafsika H Georgopapadakou
Jul 19, 2006·Progress in Lipid Research·Ponniah SelvakumarRajendra K Sharma
Jan 13, 2005·Genome Génome / Conseil National De Recherches Canada·Tim DumonceauxRaju Datla
Mar 29, 2000·The Journal of Biological Chemistry·Q QiR Datla
Sep 25, 1998·The Journal of Biological Chemistry·K AshrafiJ I Gordon
May 22, 2007·The Journal of Biological Chemistry·Jian WuJianping Ding
Jun 20, 1998·The Journal of Biological Chemistry·J K LodgeJ I Gordon
Dec 19, 2002·The Journal of Biological Chemistry·Helen P PriceDeborah F Smith
Aug 19, 2010·The FEBS Journal·Seiji YamauchiYuzuru Tozawa
Dec 9, 1997·Archives of Biochemistry and Biophysics·R V RajuR K Sharma
Jun 20, 1998·Experimental Cell Research·R V RajuR K Sharma
Jun 10, 1998·Microbiology and Molecular Biology Reviews : MMBR·P StanleyC Hughes