Phosphatase activity characterization on the surface of intact bloodstream forms of Trypanosoma brucei
Procyclic forms of Trypanosoma brucei possess a phosphatase activity on their external cell surface. This activity, while it dephosphorylates [32P]phosphocasein, is inhibited weakly by NaF and tartrate but strongly by vanadate. In this work, we describe the presence of an external phosphatase activi...
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Published in | FEMS microbiology letters Vol. 220; no. 2; pp. 197 - 206 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
Oxford
Oxford University Press
01.03.2003
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Abstract | Procyclic forms of Trypanosoma brucei possess a phosphatase activity on their external cell surface. This activity, while it dephosphorylates [32P]phosphocasein, is inhibited weakly by NaF and tartrate but strongly by vanadate. In this work, we describe the presence of an external phosphatase activity in intact bloodstream forms of T. brucei. With p-nitrophenyl phosphate (pNPP) as substrate, these intact cells produced 3–5 nmol pNP min−1 mg−1, linearly for up to at least 30 min. The activity was not significantly increased by Mg2+, Mn2+, Ca2+ and Co2+, but was inhibited by vanadate, NaF, p-chloromercuribenzoate and Zn2+ and was insensitive to okadaic acid. Membrane-enriched fractions of parasites contained an acid phosphatase activity, with a pH optimum in the range of 4.5–5.5. This activity hydrolyzed phosphotyrosine (40 nmol phosphate min−1 mg−1) better than phosphothreonine or phosphoserine. Partial purification of this phosphatase yielded a single activity band following gel electrophoresis, a Km value of 0.29 mM with pNPP and was insensitive to the Fe2+/H2O2/ascorbate system. |
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AbstractList | Procyclic forms of Trypanosoma brucei possess a phosphatase activity on their external cell surface. This activity, while it dephosphorylates [32P]phosphocasein, is inhibited weakly by NaF and tartrate but strongly by vanadate. In this work, we describe the presence of an external phosphatase activity in intact bloodstream forms of T. brucei. With p-nitrophenyl phosphate (pNPP) as substrate, these intact cells produced 3–5 nmol pNP min−1 mg−1, linearly for up to at least 30 min. The activity was not significantly increased by Mg2+, Mn2+, Ca2+ and Co2+, but was inhibited by vanadate, NaF, p-chloromercuribenzoate and Zn2+ and was insensitive to okadaic acid. Membrane-enriched fractions of parasites contained an acid phosphatase activity, with a pH optimum in the range of 4.5–5.5. This activity hydrolyzed phosphotyrosine (40 nmol phosphate min−1 mg−1) better than phosphothreonine or phosphoserine. Partial purification of this phosphatase yielded a single activity band following gel electrophoresis, a Km value of 0.29 mM with pNPP and was insensitive to the Fe2+/H2O2/ascorbate system. |
Author | José Mauro Granjeiro Meyer-Fernandes, José Roberto Aoyama, Hiroshi Eulázio Mikio Taga Fernandes, Eloise Cedro |
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Copyright | 2003 Federation of European Microbiological Societies |
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Snippet | Procyclic forms of Trypanosoma brucei possess a phosphatase activity on their external cell surface. This activity, while it dephosphorylates... |
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SubjectTerms | Acid phosphatase Ascorbic acid Calcium Calcium ions Carbon dioxide Cell surface Cobalt Electrophoresis Gel electrophoresis Hydrogen peroxide Iron Kinases Magnesium Microbiology Okadaic acid p-Nitrophenylphosphate Parasites Phosphatase Phosphoserine Phosphotyrosine Protozoa Purification Substrates Trypanosoma brucei Vanadate Vanadates Zinc |
Title | Phosphatase activity characterization on the surface of intact bloodstream forms of Trypanosoma brucei |
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