caa a22 4500 477119786 CHVBK 20180405111633.0 cr unu---uuuuu 170330e19960301xx s 000 0 eng 10.1007/BF00714335 doi (NATIONALLICENCE)springer-10.1007/BF00714335 Expression of human N-myristoyltransferase in Escherichia coli [Elektronische Daten] Comparison with N-myristoyltransferases expressed in different tissues [Rajala Raju, Raju Datla, Rajendra Sharma] Myristoyl CoA:protein N-myristoyltransferase catalyzes the addition of myristate to the amino-terminal glycine residue of a number of eukaryotic proteins.Escherichia coli transformed with human NMT expression construct produced high levels of N-myristoyltransferase. Using the combination of ammonium sulfate precipitation, chromatography on SP-Sepharose fast flow and fast protein liquid chromatography on Mono-S, the enzyme was purified more than 100 fold with 40% yield. The hNMT fusion protein exhibited an apparent molecular weight of 53 kDa on SDS-polyacrylamide gel electrophoresis. Upon cleavage by the Enterokinase [(Asp)4-Lys], the hNMT exhibited an apparent molecular mass of 49 kDa without loss of catalytic activity. The hNMT activity could be greatly activated severalfold with the use of Tris, SDS, ethanol and acetonitrile. The catalytic activity of hNMT was potently inhibited in a concentration dependent manner by NIP711 a bovine brain NMT inhibitory protein with a half maximal inhibition of 31.0 nM. TheE. coli expressed hNMT was homogeneous and showed enzyme activity. Kluwer Academic Publishers, 1996 myristoylation nationallicence myristoyltransferase nationallicence myristoyl CoA nationallicence inhibition nationallicence activation nationallicence purification nationallicence NMT : N-myristoyl CoA:protein N-myristoyltransferase nationallicence NIP71 : 71 kDa heat stable membrane bound N-myristoyltransferase inhibitor protein nationallicence hNMT : human NMT nationallicence DTNB : N-5,5′dithiobis (2-nitrobenzoic acid) nationallicence FPLC : fast protein liquid chromatography nationallicence IPTG : isopropyl β-D-thiogalactopyranoside nationallicence cDNA : complementarydeoxyribonucleic acid nationallicence SDS : sodium dodecyl sulfate nationallicence PMSF : phenylmethylsulfonyl fluoride nationallicence Raju Rajala Department of Pathology and Saskatoon Cancer Centre, College of Medicine, University of Saskatchewan, Royal University Hospital, S7N 4H4, Saskatoon, Saskatchewan aut Datla Raju Plant Biotechnology Institute, National Research Council, 110 Gymnasium place, S7N OW9, Saskatchewan, Canada aut Sharma Rajendra Department of Pathology and Saskatoon Cancer Centre, College of Medicine, University of Saskatchewan, Royal University Hospital, S7N 4H4, Saskatoon, Saskatchewan aut Molecular and Cellular Biochemistry Kluwer Academic Publishers 155/1(1996-03-01), 69-76 0300-8177 155:1<69 1996 155 11010 https://doi.org/10.1007/BF00714335 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00714335 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Raju Rajala Department of Pathology and Saskatoon Cancer Centre, College of Medicine, University of Saskatchewan, Royal University Hospital, S7N 4H4, Saskatoon, Saskatchewan aut NATIONALLICENCE 700 1- Datla Raju Plant Biotechnology Institute, National Research Council, 110 Gymnasium place, S7N OW9, Saskatchewan, Canada aut NATIONALLICENCE 700 1- Sharma Rajendra Department of Pathology and Saskatoon Cancer Centre, College of Medicine, University of Saskatchewan, Royal University Hospital, S7N 4H4, Saskatoon, Saskatchewan aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Kluwer Academic Publishers 155/1(1996-03-01), 69-76 0300-8177 155:1<69 1996 155 11010 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer