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Title: Enzymatic and thermodynamic profiles of a heterotetramer lactate dehydrogenase isozyme in swine

Abstract

Lactate dehydrogenase (LDH) is a glycolytic enzyme that catalyzes the final step of glycolysis and produces NAD{sup +}. In somatic cells, LDH forms homotetramers and heterotetramers that are encoded by two different genes: LDHA (skeletal muscle type, M) and LDHB (heart type, H). Analysis of LDH isozymes is important for understanding the physiological role of homotetramers and heterotetramers and for optimizing inhibition of their enzymatic activity as it may result in distinct effects. Previously, we reported that hydroxychloroquine (HCQ) inhibited LDH activity, but we did not examine isozyme specificity. In the present study, we isolated heterotetrameric LDH (H{sub 2}M{sub 2}) from swine brain, determined its kinetic and thermodynamic properties, and examined the effect of HCQ on its activity compared to homotetrameric LDH isozymes. We show that: (1) the K{sub m} values for H{sub 2}M{sub 2}–mediated catalysis of pyruvate or lactate were intermediate compared to those for the homotetrameric isozymes, M{sub 4} and H{sub 4} whereas the V{sub max} values were similar; (2) the K{sub m} and V{sub max} values for H{sub 2}M{sub 2}–mediated catalysis of NADH were not significantly different among LDH isozymes; (3) the values for activation energy and van't Hoff enthalpy changes for pyruvate reduction of H{sub 2}M{submore » 2} were intermediate compared to those for the homotetrameric isozymes; (4) the temperature for half residual activity of H{sub 2}M{sub 2} was closer to that for M{sub 4} than for H{sub 4}. We also show that HCQ had different affinities for various LDH isozymes. - Highlights: • Heterotetrameric (H{sub 2}M{sub 2}) LDH isozyme was isolated from swine brain. • Kinetics of H{sub 2}M{sub 2} were intermediate between the two homotetramers. • Thermodynamics of H{sub 2}M{sub 2} were also intermediate between the two homotetramers. • Hydroxychloroquine inhibited more strongly H{sub 2}M{sub 2} than homotetramers.« less

Authors:
 [1];  [2];  [3];  [4];  [1];  [1]
  1. Department of Life Science, Graduate School of Engineering Science, Akita University (Japan)
  2. Department of Hematology, Nephrology, and Rheumatology, Master Course of Graduate School of Medicine, Akita University (Japan)
  3. Department of Creative Engineering, Kitakyusyu National College of Technology (Japan)
  4. Department of Bioengineering, Graduate School of Engineering, Nagaôka University of Technology (Japan)
Publication Date:
OSTI Identifier:
22696670
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 479; Journal Issue: 4; Other Information: Copyright (c) 2016 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ACTIVATION ENERGY; INHIBITION; LACTATE DEHYDROGENASE; SOCIO-ECONOMIC FACTORS; SOMATIC CELLS; SWINE; THERMODYNAMICS

Citation Formats

Goto, Tatsufumi, Sugawara, Kotomi, Nakamura, Shigeyoshi, Kidokoro, Shun-Ichi, Wakui, Hideki, Nunomura, Wataru, and Research Center for Engineering Science, Graduate School of Engineering Science, Akita University. Enzymatic and thermodynamic profiles of a heterotetramer lactate dehydrogenase isozyme in swine. United States: N. p., 2016. Web. doi:10.1016/J.BBRC.2016.09.118.
Goto, Tatsufumi, Sugawara, Kotomi, Nakamura, Shigeyoshi, Kidokoro, Shun-Ichi, Wakui, Hideki, Nunomura, Wataru, & Research Center for Engineering Science, Graduate School of Engineering Science, Akita University. Enzymatic and thermodynamic profiles of a heterotetramer lactate dehydrogenase isozyme in swine. United States. doi:10.1016/J.BBRC.2016.09.118.
Goto, Tatsufumi, Sugawara, Kotomi, Nakamura, Shigeyoshi, Kidokoro, Shun-Ichi, Wakui, Hideki, Nunomura, Wataru, and Research Center for Engineering Science, Graduate School of Engineering Science, Akita University. Fri . "Enzymatic and thermodynamic profiles of a heterotetramer lactate dehydrogenase isozyme in swine". United States. doi:10.1016/J.BBRC.2016.09.118.
@article{osti_22696670,
title = {Enzymatic and thermodynamic profiles of a heterotetramer lactate dehydrogenase isozyme in swine},
author = {Goto, Tatsufumi and Sugawara, Kotomi and Nakamura, Shigeyoshi and Kidokoro, Shun-Ichi and Wakui, Hideki and Nunomura, Wataru and Research Center for Engineering Science, Graduate School of Engineering Science, Akita University},
abstractNote = {Lactate dehydrogenase (LDH) is a glycolytic enzyme that catalyzes the final step of glycolysis and produces NAD{sup +}. In somatic cells, LDH forms homotetramers and heterotetramers that are encoded by two different genes: LDHA (skeletal muscle type, M) and LDHB (heart type, H). Analysis of LDH isozymes is important for understanding the physiological role of homotetramers and heterotetramers and for optimizing inhibition of their enzymatic activity as it may result in distinct effects. Previously, we reported that hydroxychloroquine (HCQ) inhibited LDH activity, but we did not examine isozyme specificity. In the present study, we isolated heterotetrameric LDH (H{sub 2}M{sub 2}) from swine brain, determined its kinetic and thermodynamic properties, and examined the effect of HCQ on its activity compared to homotetrameric LDH isozymes. We show that: (1) the K{sub m} values for H{sub 2}M{sub 2}–mediated catalysis of pyruvate or lactate were intermediate compared to those for the homotetrameric isozymes, M{sub 4} and H{sub 4} whereas the V{sub max} values were similar; (2) the K{sub m} and V{sub max} values for H{sub 2}M{sub 2}–mediated catalysis of NADH were not significantly different among LDH isozymes; (3) the values for activation energy and van't Hoff enthalpy changes for pyruvate reduction of H{sub 2}M{sub 2} were intermediate compared to those for the homotetrameric isozymes; (4) the temperature for half residual activity of H{sub 2}M{sub 2} was closer to that for M{sub 4} than for H{sub 4}. We also show that HCQ had different affinities for various LDH isozymes. - Highlights: • Heterotetrameric (H{sub 2}M{sub 2}) LDH isozyme was isolated from swine brain. • Kinetics of H{sub 2}M{sub 2} were intermediate between the two homotetramers. • Thermodynamics of H{sub 2}M{sub 2} were also intermediate between the two homotetramers. • Hydroxychloroquine inhibited more strongly H{sub 2}M{sub 2} than homotetramers.},
doi = {10.1016/J.BBRC.2016.09.118},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 4,
volume = 479,
place = {United States},
year = {2016},
month = {10}
}