Effect of Borate on the Catalytic Activities of Muscle Glyceraldehyde‐3‐Phosphate Dehydrogenase

Wolny, M

doi:10.1111/j.1432-1033.1977.tb11911.x

Cited by 14 publications

(2 citation statements)

References 26 publications

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“…This enzyme is broadly distributed in nature in a variety of organisms ranging from bacterial cells to human tissues [4,5]. It is found for the most part in the cytosol, mitochondria, and chloroplasts in plants.…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of glyceraldehyde-3-phosphate dehydrogenase from saline strain <i>Idiomarina loihiensis</i>

Mardad¹,

Baibai²,

Ammar³

et al. 2013

ABC

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Idiomarina loihiensis was isolated from the salt works in Sfax (Tunisia), until now, the characterization of the GAPDH phosphorylante was never studied. Here, we report the isolation and the biochemical characterization of glyceralehyde-3-phosphate dehydrogenase (GAPDH) from I. loihiensis saline's bacteria on the basis of the apparent native and subunit molecular weights, physico-chemical and kinetic characterizations. The purification method consisted of two steps, ammonium sulfate fractionation followed by one chromatographic step, namely dye-affinity on Blue Sepharose CL-6B. Polyclonal antibodies against the purified enzyme were used to recognize the I. loihiensis GAPDH by Western blotting. The optimum pH of the purified enzyme was 8.5. Studies on the effect of temperatures revealed an enzyme increasing activity of about 45˚C. The molecular weight of the purified enzyme was 36 kDa determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Non-denaturing polyacrylamide gels yield a molecular weight of 147 kDa. The Michaelis constants for NAD + and D-glyceraldehyde-3-phosphate estimated was 19 μM and 3.1 μM, respectively. The maximal velocity of the purified enzyme was estimated to be 2.06 U/mg, approximately 6-fold increase in specific activity and a final yield of approximately 32.5%. The physicochemical properties of this GAPDH, being characterized, could be used in further studies.

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Section: Introductionmentioning

confidence: 99%

Purification and characterization of glyceraldehyde-3-phosphate dehydrogenase from saline strain <i>Idiomarina loihiensis</i>

Mardad¹,

Baibai²,

Ammar³

et al. 2013

ABC

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“…Glycolysis would be the logical metabolic site of action. Borate is known to competitively and reversibly inhibit the activity of purified glyceraldehyde-3-phosphate dehydrogenase through boration of the cofactor nicotinamide adenine dinucleotide (NAD) (36). Consequently, it is possible that this or other NAD-requiring enzymes may be affected.…”

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Mechanism of the testicular toxicity of boric acid in rats: in vivo and in vitro studies.

Chapin²

1994

Environ Health Perspect

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High-dose boric acid (BA) exposure produces testicular lesions in adult rats characterized by inhibited spermiation (IS) that may progress to atrophy. In vivo and in vitro studies addressed possible mechanisms. In vivo, boron tissue disposition was examined, since no detailed data existed, and relevant boron concentrations for in vitro studies needed to be set. Since BA induces riboflavinuria and also affects calcium/phosphorus homeostasis, and testis zinc appears essential for normal testis function, we examined BA effects on flavin status and testis levels of phosphorus (P), calcium (Ca) and zinc (Zn). Data showed that the testicular toxicity and central nervous system (CNS) hormonal effect were not due to selective boron accumulation in testis or brain/hypothalamus, with testis boron concentrations at approximately 1 to 2 mM; that riboflavin deficiency is not involved, due to both the absence of overt signs of deficiency and effects on tissue flavin content during BA exposure; and that changes in testis P, Ca and Zn levels did not precede atrophy, and are therefore unlikely to be mechanistically relevant. In vitro studies addressed the hallmarks of the BA testicular toxicity: the mild hormone effect, the initial IS, and atrophy. No effect of BA on the steroidogenic function of isolated Leydig cells was observed, supporting the contention of a CNS-mediated rather than a direct hormone effect. Since increased testicular cyclic adenosine monophosphate (cAMP) produces IS, and a role for the serine proteases plasminogen activators (PAs) in spermiation has been proposed, we examined in vitro BA effects on both Sertoli cell cAMP accumulation and PA activity, respectively. Results showed that the IS is not due to BA effects on either process. To address the atrophy, we evaluated BA effects in Sertoli-germ cell cocultures on: Sertoli cell energy metabolism, since lactate, secreted by Sertoli cells, is a preferred energy source for germ cells; and also on DNA/RNA synthesis, since germ cells synthesize DNA/RNA and BA impairs nucleic acid synthesis in liver, and may do so in testis. The most sensitive in vitro endpoint was DNA synthesis of mitotic/meiotic germ cells, with energy metabolism in Sertoli or germ cells affected to a lesser extent. The in vitro effect on DNA synthesis was manifest in vivo as a decrease in the early germ cell/Sertoli cell ratio prior to atrophy in testes from BA-exposed rats. Overall, these combined studies revealed some changes offering a plausible explanation for the atrophy aspect of the BA testicular lesion. However, the mechanism for the IS is still undefined, and should be the subject of future work. -Environ Health Perspect 102(Suppl 7): 99-105 (1994)

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Borsäure [MAK Value Documentation in German language, 1991]

2012

The MAK‐Collection for Occupational Health and Safety

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Veröffentlicht in der Reihe Gesundheitsschädliche Arbeitsstoffe , 17. Lieferung, Ausgabe 1991 Der Artikel enthält folgende Kapitel: Besondere Hinweise Allgemeiner Wirkungscharakter Angaben zur Pharmakokinetik Biologische Wirkungen Erfahrungen beim Menschen Borsäure‐Vergütungen und Vergiftungssymptome Akute toxische Wirkungen Wirkung nach Haut‐ und Schleimhaut‐Applikation Subchronische und chronische Wirkungen Tierexperimentelle Befunde Akute toxische Wirkungen Wirkung am Auge sowie nach Haut‐ und Schleimhaut‐Applikation Subchronische und chronische Wirkungen Lokale Wirkung auf Haut und Schleimhäute Allergene Wirkung Reproduktions‐ und Teratogenitätsstudien Untersuchungen zur gentoxischen Wirkung Untersuchungen zur kanzerogenen Wirkung Bewertung

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Effect of Borate on the Catalytic Activities of Muscle Glyceraldehyde‐3‐Phosphate Dehydrogenase

Cited by 14 publications

References 26 publications

Purification and characterization of glyceraldehyde-3-phosphate dehydrogenase from saline strain <i>Idiomarina loihiensis</i>

Purification and characterization of glyceraldehyde-3-phosphate dehydrogenase from saline strain <i>Idiomarina loihiensis</i>

Mechanism of the testicular toxicity of boric acid in rats: in vivo and in vitro studies.

Borsäure [MAK Value Documentation in German language, 1991]

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Effect of Borate on the Catalytic Activities of Muscle Glyceraldehyde‐3‐Phosphate Dehydrogenase

Cited by 14 publications

References 26 publications

Purification and characterization of glyceraldehyde-3-phosphate dehydrogenase from saline strain &lt;i&gt;Idiomarina loihiensis&lt;/i&gt;

Purification and characterization of glyceraldehyde-3-phosphate dehydrogenase from saline strain &lt;i&gt;Idiomarina loihiensis&lt;/i&gt;

Mechanism of the testicular toxicity of boric acid in rats: in vivo and in vitro studies.

Borsäure [MAK Value Documentation in German language, 1991]

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Purification and characterization of glyceraldehyde-3-phosphate dehydrogenase from saline strain <i>Idiomarina loihiensis</i>

Purification and characterization of glyceraldehyde-3-phosphate dehydrogenase from saline strain <i>Idiomarina loihiensis</i>