Simultaneous purification of L-malate dehydrogenase and L-lactate dehydrogenase from bovine heart by biomimetic-dye affinity chromatography

Labrou, Nikolaos E.; Clonis, Yannis D.

doi:10.1007/s004490050303

Cited by 6 publications

(3 citation statements)

References 25 publications

(53 reference statements)

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“…LDH Purification and the Activity Assay In Vitro LDH was extracted and purified from heart of control mice according to Labrou and Clonis method [15]. In order to further investigate the effects of nanoparticulate anatase TiO 2 on LDH activity, the in vitro purified LDH activity assays were performed.…”

Section: Ldh Activity Assay In Vivomentioning

confidence: 99%

Interaction Between Nanoparticulate Anatase TiO2 and Lactate Dehydrogenase

Duan

Liu

et al. 2009

Biol Trace Elem Res

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In order to study the mechanisms underlying the effects of TiO(2) nanoparticles on lactate dehydrogenase (LDH, EC1.1.1.27), Institute of Cancer Research region mice were injected with nanoparticulate anatase TiO(2) (5 nm) of various doses into the abdominal cavity daily for 14 days. We then examined LDH activity in vivo and in vitro and direct evident for interaction between nanoparticulate anatase TiO(2) and LDH using spectral methods. The results showed that nanoparticulate anatase TiO(2) could significantly activate LDH in vivo and in vitro; the kinetics constant (Km) and Vmax were 0.006 microM and 1,149 unit mg(-1) protein min(-1), respectively, at a low concentration of nanoparticulate anatase TiO(2), and 3.45 and 0.031 microM and 221 unit mg(-1) protein min(-1), respectively, at a high concentration of nanoparticulate anatase TiO(2). By fluorescence spectral assays, the nanoparticulate anatase TiO(2) was determined to be directly bound to LDH, and the binding constants of the binding site were 1.77 x 10(8) L mol(-1) and 2.15 x 10(7) L mol(-1), respectively, and the binding distance between nanoparticulate anatase TiO(2) and the Trp residue of LDH was 4.18 nm, and nanoparticulate anatase TiO(2) induced the protein unfolding. It was concluded that the binding of nanoparticulate anatase TiO(2) altered LDH structure and function.

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Section: Ldh Activity Assay In Vivomentioning

confidence: 99%

Interaction Between Nanoparticulate Anatase TiO2 and Lactate Dehydrogenase

Duan

Liu

et al. 2009

Biol Trace Elem Res

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“…LDH was extracted and purified from heart of control mice according to Labrou and Clonis method [25]. Frozen tissue was cut into small pieces and frozen again at −20°C (15 g).…”

Section: Ldh Purification and The Activity Assay In Vitromentioning

confidence: 99%

“…The experimental groups were given LaCl 3 , CeCl 3 , and NdCl 3 at a dose of 2. 5,5,10,15,20,25, and 30 mg/kg BW daily by intraperitoneal injection for 14 days, respectively. The symptom and mortality were observed and recorded carefully every day.…”

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confidence: 99%

The Effects of Lanthanoid on the Structure–Function of Lactate Dehydrogenase from Mice Heart

Duan

Zhou

et al. 2009

Biol Trace Elem Res

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The activity of lactate dehydrogenase (LDH, EC1.1.1.27) is often changed upon inflammatory responses in animals. Lanthanoid (Ln) was shown to provoke various inflammatory responses both in rats and mice; however, the molecular mechanism by which Ln3+ exert its toxicity has not been completely understood, especially that we know little about the mechanism of the interaction between Ln with 4f electron shell and alternation valence and LDH. In this report, we investigated the mechanisms of LaCl3, CeCl3, and NdCl3 on LDH activity in vivo and in vitro. Our results showed that La3+, Ce3+, and Nd3+ could significantly activate LDH in vivo and in vitro; the order of activation was Ce3+>Nd3+> La3+>control. The affinity of LDH for Ce3+ was higher than Nd3+ and La3+; the saturated binding sites for Ce3+ on the LDH protein were 1.2 and for La3+ and Nd3+ 1.55. Ln3+ caused the reduction of exposure degree of cysteine or tryptophan/tyrosine of LDH, the increase of space resistance, and the enhancement of α-helix in secondary structure of LDH, which was greatest in Ce3+ treatment, medium in Nd3+ treatment, and least in La3+ treatment. It implied that the changes of structure-function on LDH caused by Ln3+ were closely related to the characteristics of 4f electron shell and alternation valence in Ln.

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