Phosphorylation of Ser-204 and Tyr-405 in human malonyl-CoA decarboxylase expressed in silkworm Bombyx mori regulates catalytic decarboxylase activity
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| 008 | 210128e20151101xx s 000 0 eng | ||
| 024 | 7 | 0 | |a 10.1007/s00253-015-6687-x |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6687-x | ||
| 245 | 0 | 0 | |a Phosphorylation of Ser-204 and Tyr-405 in human malonyl-CoA decarboxylase expressed in silkworm Bombyx mori regulates catalytic decarboxylase activity |h [Elektronische Daten] |c [In-Wook Hwang, Yu Makishima, Tomohiro Suzuki, Tatsuya Kato, Sungjo Park, Andre Terzic, Shin-kyo Chung, Enoch Park] |
| 520 | 3 | |a Decarboxylation of malonyl-CoA to acetyl-CoA by malonyl-CoA decarboxylase (MCD; EC 4.1.1.9) is a vital catalytic reaction of lipid metabolism. While it is established that phosphorylation of MCD modulates the enzymatic activity, the specific phosphorylation sites associated with the catalytic function have not been documented due to lack of sufficient production of MCD with proper post-translational modifications. Here, we used the silkworm-based Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid system to express human MCD (hMCD) and mapped phosphorylation effects on enzymatic function. Purified MCD from silkworm displayed post-translational phosphorylation and demonstrated coherent enzymatic activity with high yield (−200μg/silkworm). Point mutations in putative phosphorylation sites, Ser-204 or Tyr-405 of hMCD, identified by bioinformatics and proteomics analyses reduced the catalytic activity, underscoring the functional significance of phosphorylation in modulating decarboxylase-based catalysis. Identified phosphorylated residues are distinct from the decarboxylation catalytic site, implicating a phosphorylation-induced global conformational change of MCD as responsible in altering catalytic function. We conclude that phosphorylation of Ser-204 and Tyr-405 regulates the decarboxylase function of hMCD leveraging the silkworm-based BmNPV bacmid expression system that offers a fail-safe eukaryotic production platform implementing proper post-translational modification such as phosphorylation. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a Human malonyl-CoA decarboxylase (hMCD) |2 nationallicence | |
| 690 | 7 | |a Site-directed mutagenesis |2 nationallicence | |
| 690 | 7 | |a Phosphorylation/dephosphorylation |2 nationallicence | |
| 690 | 7 | |a Lipid metabolism |2 nationallicence | |
| 690 | 7 | |a Silkworm |2 nationallicence | |
| 690 | 7 | |a Bombyx mori nucleopolyhedrovirus |2 nationallicence | |
| 700 | 1 | |a Hwang |D In-Wook |u Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | |
| 700 | 1 | |a Makishima |D Yu |u Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | |
| 700 | 1 | |a Suzuki |D Tomohiro |u Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | |
| 700 | 1 | |a Kato |D Tatsuya |u Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | |
| 700 | 1 | |a Park |D Sungjo |u Center for Regenerative Medicine, Mayo Clinic, 200 First Street SW, 55905, Rochester, MN, USA |4 aut | |
| 700 | 1 | |a Terzic |D Andre |u Center for Regenerative Medicine, Mayo Clinic, 200 First Street SW, 55905, Rochester, MN, USA |4 aut | |
| 700 | 1 | |a Chung |D Shin-kyo |u School of Food Science and Biotechnology, Kyungpook National University, 702-701, Daegu, Republic of Korea |4 aut | |
| 700 | 1 | |a Park |D Enoch |u Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/21(2015-11-01), 8977-8986 |x 0175-7598 |q 99:21<8977 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6687-x |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-015-6687-x |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Hwang |D In-Wook |u Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Makishima |D Yu |u Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Suzuki |D Tomohiro |u Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Kato |D Tatsuya |u Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Park |D Sungjo |u Center for Regenerative Medicine, Mayo Clinic, 200 First Street SW, 55905, Rochester, MN, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Terzic |D Andre |u Center for Regenerative Medicine, Mayo Clinic, 200 First Street SW, 55905, Rochester, MN, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Chung |D Shin-kyo |u School of Food Science and Biotechnology, Kyungpook National University, 702-701, Daegu, Republic of Korea |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Park |D Enoch |u Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/21(2015-11-01), 8977-8986 |x 0175-7598 |q 99:21<8977 |1 2015 |2 99 |o 253 | ||