Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme
| ISSN: |
0167-4838
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|---|---|
| Keywords: |
(Human skeletal muscle) ; (Muscle mitrochondrion) ; ATP affinity chromatography ; Malic enzyme ; NADP^+ ; NAD^+
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| Source: |
Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
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| Topics: |
Biology
Chemistry and Pharmacology
Medicine
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| Type of Medium: |
Electronic Resource
|
| URL: |
| _version_ | 1798291642216611840 |
|---|---|
| autor | Taroni, F. Gellera, C. Di Donato, S. |
| autorsonst | Taroni, F. Gellera, C. Di Donato, S. |
| book_url | http://linkinghub.elsevier.com/retrieve/pii/0167-4838(87)90191-9 |
| datenlieferant | nat_lic_papers |
| fussnote | Human muscle mitochondria reduced either NADP^+ or NAD^+ in the presence of l-malate and Mn^2^+ or Mg^2^+. After polyacrylamide slab gel electrophoresis and agarose gel isoelectrofocusing, two bands were seen in mitochondrial extract, one strictly NADP^+-dependent and the other reacting with either NAD^+ or NADP^+. The two mitochondrial malic enzymes were separated by DEAE-Sepharose chromatography. The NAD^+/NADP^+-dependent enzyme was purified 1600-fold with a final yield of 34% and a final specific activity of 32.9 units/mg of protein by employing affinity chromatography on Agarose-ATP. SDS electrophoresis revealed a single band having an apparent M"r = 64 000. Estimates of the native apparent molecular weight upon gel filtration yielded a value of 140 300. Kinetic characterization showed that succinate and ATP were activator and inhibitor, respectively. In the absence of succinate the K"m values for malate, NAD^+ and NADP^+ were 3.7, 0.13 and 0.78 mM, respectively; in the presence of succinate the K"m value for malate was 1.9 mM. ATP was found to be an inhibitor competitive with malate, with a K"i (ATP) of 0.2 mM. This is the first report to show that mammalian skeletal muscle mitochondria contains two distinct malic enzymes, one active with either NAD^+ or NADP^+ and the other active only with NADP^+. |
| hauptsatz | hsatz_simple |
| identnr | NLZ186890869 |
| issn | 0167-4838 |
| journal_name | Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular |
| materialart | 1 |
| package_name | Elsevier |
| publikationsort | Amsterdam |
| publisher | Elsevier |
| reference | 916 (1987), S. 446-454 |
| schlagwort | (Human skeletal muscle) (Muscle mitrochondrion) ATP affinity chromatography Malic enzyme NADP^+ NAD^+ |
| search_space | articles |
| shingle_author_1 | Taroni, F. Gellera, C. Di Donato, S. |
| shingle_author_2 | Taroni, F. Gellera, C. Di Donato, S. |
| shingle_author_3 | Taroni, F. Gellera, C. Di Donato, S. |
| shingle_author_4 | Taroni, F. Gellera, C. Di Donato, S. |
| shingle_catch_all_1 | Taroni, F. Gellera, C. Di Donato, S. Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme (Human skeletal muscle) (Muscle mitrochondrion) ATP affinity chromatography Malic enzyme NADP^+ NAD^+ (Human skeletal muscle) (Muscle mitrochondrion) ATP affinity chromatography Malic enzyme NADP^+ NAD^+ 0167-4838 01674838 Elsevier |
| shingle_catch_all_2 | Taroni, F. Gellera, C. Di Donato, S. Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme (Human skeletal muscle) (Muscle mitrochondrion) ATP affinity chromatography Malic enzyme NADP^+ NAD^+ (Human skeletal muscle) (Muscle mitrochondrion) ATP affinity chromatography Malic enzyme NADP^+ NAD^+ 0167-4838 01674838 Elsevier |
| shingle_catch_all_3 | Taroni, F. Gellera, C. Di Donato, S. Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme (Human skeletal muscle) (Muscle mitrochondrion) ATP affinity chromatography Malic enzyme NADP^+ NAD^+ (Human skeletal muscle) (Muscle mitrochondrion) ATP affinity chromatography Malic enzyme NADP^+ NAD^+ 0167-4838 01674838 Elsevier |
| shingle_catch_all_4 | Taroni, F. Gellera, C. Di Donato, S. Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme (Human skeletal muscle) (Muscle mitrochondrion) ATP affinity chromatography Malic enzyme NADP^+ NAD^+ (Human skeletal muscle) (Muscle mitrochondrion) ATP affinity chromatography Malic enzyme NADP^+ NAD^+ 0167-4838 01674838 Elsevier |
| shingle_title_1 | Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme |
| shingle_title_2 | Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme |
| shingle_title_3 | Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme |
| shingle_title_4 | Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme |
| sigel_instance_filter | dkfz geomar wilbert ipn albert fhp |
| source_archive | Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 |
| timestamp | 2024-05-06T08:35:52.456Z |
| titel | Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme |
| titel_suche | Evidence for two distinct mitochondrial malic enzymes in human skeletal muscle: purification and properties of the NAD(P)^+-dependent enzyme Human muscle mitochondria reduced either NADP^+ or NAD^+ in the presence of l-malate and Mn^2^+ or Mg^2^+. After polyacrylamide slab gel electrophoresis and agarose gel isoelectrofocusing, two bands were seen in mitochondrial extract, one strictly NADP^+-dependent and the other reacting with either NAD^+ or NADP^+. The two mitochondrial malic enzymes were separated by DEAE-Sepharose chromatography. The NAD^+/NADP^+-dependent enzyme was purified 1600-fold with a final yield of 34% and a final specific activity of 32.9 units/mg of protein by employing affinity chromatography on Agarose-ATP. SDS electrophoresis revealed a single band having an apparent M"r = 64 000. Estimates of the native apparent molecular weight upon gel filtration yielded a value of 140 300. Kinetic characterization showed that succinate and ATP were activator and inhibitor, respectively. In the absence of succinate the K"m values for malate, NAD^+ and NADP^+ were 3.7, 0.13 and 0.78 mM, respectively; in the presence of succinate the K"m value for malate was 1.9 mM. ATP was found to be an inhibitor competitive with malate, with a K"i (ATP) of 0.2 mM. This is the first report to show that mammalian skeletal muscle mitochondria contains two distinct malic enzymes, one active with either NAD^+ or NADP^+ and the other active only with NADP^+. |
| topic | W V WW-YZ |
| uid | nat_lic_papers_NLZ186890869 |