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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
5-6
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pubmed:dateCreated |
1982-5-21
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pubmed:abstractText |
Lactate dehydrogenase (LDH) from Plasmodium falciparum was partially purified by two different procedures. In the first procedure, parasitized erythrocytes (80% parasitemia) were lysed, and the soluble fraction was purified on DEAE-Sephadex to separate the parasite LDH(LDH-P) from the LDH isoenzymes present in the human erythrocytes. LDH-P was then purified by high-performance liquid chromatography (HPLC) on a TSK-G-3000 SW protein column. This two-step procedure gave LDH-P with specific activity 85 micromol/min/mg protein; this represented a 700-fold increase in specific activity relative to the starting lysate. Alternatively, parasites of P. falciparum were isolated by mechanical rupture of infected erythrocytes followed by differential centrifugation. The 100,000 X g supernatant obtained after lysis of these parasites showed LDH-P specific activity 3.6 micromol/min/mg protein. This activity was free of contaminating erythrocyte LDH as determined by electrophoresis and specific staining for LDH. Further purification of LDH-P by HPLC, as before, gave material with specific activity 98 micromol/min/mg protein. Recoveries of activity on HPLC were more than 90%, demonstrating the usefulness of this procedure for the partial purification of small quantities of parasite protein. The kinetic properties of LDH-P were compared with those of two of the human isozymes, LDH-H4 and LDH-M4 . LDH-P resembles LDH-H4 in its kinetic properties: KM (NADH) is 7, 8.3 and 1.3 microM for LDH-P, LDH-H4 and LDH-M4, respectively; KM (pyruvate) is 30, 60 and 180 microM for LDH-P, LDH-H4 and LDH-M4. LDH-P differs significantly from LDH-H4 and LDH-M4 in that LDH-P is not sensitive to inhibition by high pyruvate nor sensitive to inhibition by the complex between NAD+ and pyruvate. LDH-P is inactivated within seconds by sodium deoxycholate at concentrations that do not affect LDH-H4 and slowly inactivate LDH-M4.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Deoxycholic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/L-Lactate Dehydrogenase,
http://linkedlifedata.com/resource/pubmed/chemical/NAD,
http://linkedlifedata.com/resource/pubmed/chemical/Pyruvates,
http://linkedlifedata.com/resource/pubmed/chemical/Pyruvic Acid
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pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
0166-6851
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
31
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pubmed:volume |
4
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
255-64
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:7038478-Animals,
pubmed-meshheading:7038478-Centrifugation,
pubmed-meshheading:7038478-Chromatography, High Pressure Liquid,
pubmed-meshheading:7038478-Chromatography, Ion Exchange,
pubmed-meshheading:7038478-Deoxycholic Acid,
pubmed-meshheading:7038478-Erythrocytes,
pubmed-meshheading:7038478-Kinetics,
pubmed-meshheading:7038478-L-Lactate Dehydrogenase,
pubmed-meshheading:7038478-NAD,
pubmed-meshheading:7038478-Plasmodium falciparum,
pubmed-meshheading:7038478-Pyruvates,
pubmed-meshheading:7038478-Pyruvic Acid
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pubmed:year |
1981
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pubmed:articleTitle |
Partial purification and characterization of lactate dehydrogenase from Plasmodium falciparum.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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