scholarly journals Purification of aldehyde dehydrogenase from rat liver mitochondria by α-cyanocinnamate affinity chromatography

1989 ◽  
Vol 259 (1) ◽  
pp. 105-110 ◽  
Author(s):  
R C Poole ◽  
A P Halestrap
Keyword(s):  
Molecular Mass ◽  
Rat Liver ◽  
Aldehyde Dehydrogenase ◽  
Minor Component ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Affinity Column ◽  
A Subunit ◽  
A Minor ◽  
Related Compounds

1. alpha-Cyano-4-hydroxycinnamate was coupled to Sepharose CL-4B activated with 1,2:3,4-bisepoxybutane. 2. The low-Km rat liver mitochondrial aldehyde dehydrogenase was specifically bound to this affinity medium, and could subsequently be eluted with alpha-cyano-4-hydroxycinnamate. 3. The enzyme purified in this manner had a subunit molecular mass of 55 kDa and a pI of approx. 6.5. A minor component of approx. 57 kDa was also present and had a significantly higher pI value; this may be the precursor for aldehyde dehydrogenase. 4. alpha-Cyanocinnamate and some related compounds were found to be uncompetitive inhibitors of the enzyme. 5. No cytosolic aldehyde dehydrogenase was bound to the affinity column, but a protein from a rat liver post-mitochondrial supernatant with a molecular mass of approx. 25 kDa was bound, and could be eluted subsequently with alpha-cyano-4-hydroxycinnamate.

scholarly journals Molecular cloning and characterization of a mitochondrial peroxisome proliferator-induced acyl-CoA thioesterase from rat liver

1998 ◽  
Vol 329 (3) ◽  
pp. 601-608 ◽  
Author(s):  
L. Thomas SVENSSON ◽  
T. Susanna ENGBERG ◽  
Toshifumi AOYAMA ◽  
Nobuteru USUDA ◽  
E. H. Stefan ALEXSON ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Rat Liver ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Liver Mitochondria ◽  
Leader Peptide ◽  
Rat Liver Mitochondria ◽  
Peroxisome Proliferator ◽  
Calculated Molecular Mass

We have previously reported the purification and characterization of the peroxisome proliferator-induced very-long-chain acyl-CoA thioesterase (MTE-I) from rat liver mitochondria [L. T. Svensson, S. E. H. Alexson and J. K. Hiltunen (1995) J. Biol. Chem. 270, 12177-12183]. Here we describe the cloning of the corresponding cDNA. One full-length clone was isolated that contained an open reading frame of 1359 bp encoding a polypeptide with a calculated molecular mass of 49707 Da. The deduced amino acid sequence contains a putative mitochondrial leader peptide of 42 residues. Expression of the cDNA in Chinese hamster ovary cells, followed by immunofluorescence, immunoelectron microscopy and Western blot analyses, showed that the product was targeted to mitochondria and processed to a mature protein of 45 kDa, which is similar to the molecular mass of the protein isolated from rat liver mitochondria. The recombinant enzyme showed the same acyl-CoA chain-length specificity as the isolated rat liver enzyme. Sequence analysis showed no similarity to known esterases, but a high degree (approx. 40%) of identity with bile acid-CoA:amino acid N-acyltransferase cloned from human and rat liver. A putative active-site serine motif (Gly-Xaa-Ser-Xaa-Gly) of several carboxylesterases and lipases was identified. Western and Northern blot analyses showed that MTE-I is constitutively expressed in heart and is strongly induced in liver by feeding rats with di(2-ethylhexyl)phthalate, a peroxisome proliferator, suggesting a role for the enzyme in lipid metabolism.

Dual coenzyme activities of high-Km aldehyde dehydrogenase from rat liver mitochondria

1990 ◽  
Vol 68 (4) ◽  
pp. 751-757 ◽  
Author(s):  
C. Stan Tsai ◽  
D. J. Senior
Keyword(s):  
Steady State ◽  
Rat Liver ◽  
Aldehyde Dehydrogenase ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Binary Complex ◽  
Stopped Flow ◽  
Conformation Change ◽  
Rate Limiting ◽  
Flow Burst

Various kinetic approaches were carried out to investigate kinetic attributes for the dual coenzyme activities of mitochondrial aldehyde dehydrogenase from rat liver. The enzyme catalyses NAD+- and NADP+-dependent oxidations of ethanal by an ordered bi-bi mechanism with NAD(P)+ as the first reactant bound and NAD(P)H as the last product released. The two coenzymes presumably interact with the kinetically identical site. NAD+ forms the dynamic binary complex with the enzyme, while the enzyme-NAD(P)H complex formation is associated with conformation change(s). A stopped-flow burst of NAD(P)H formation, followed by a slower steady-state turnover, suggests that either the deacylation or the release of NAD(P)H is rate limiting. Although NADP+ is reduced by a faster burst rate, NAD+ is slightly favored as the coenzyme by virtue of its marginally faster turnover rate.Key words: aldehyde dehydrogenase, coenzyme preference.

Metabolism of nitroglycerin by rat liver mitochondria and purified mitochondrial aldehyde dehydrogenase

2003 ◽  
Vol 3 (S2) ◽  
Author(s):  
Alexandra Hofer ◽  
Alexander Kollau ◽  
Wing Ming Keung ◽  
Kurt Schmidt ◽  
Bernd Mayer
Keyword(s):  
Rat Liver ◽  
Aldehyde Dehydrogenase ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria

Inactivation of aldehyde dehydrogenase in intact rat liver mitochondria by dopamine

Alcohol ◽  
1989 ◽  
Vol 6 (6) ◽  
pp. 455-460 ◽  
Author(s):  
Saffiya Catovic Turan ◽  
Pritesh Shah ◽  
Regina Pietruszko
Keyword(s):  
Rat Liver ◽  
Aldehyde Dehydrogenase ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria
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