Purification and partial characterization of alpha-mannosidase from Trypanosoma rangeli

Parasitol Res. 2000 Nov;86(11):923-8. doi: 10.1007/s004360000274.

Abstract

An alpha-mannosidase from Trypanosoma rangeli was partially purified by a protocol involving solubilization using lysis buffer followed by chromatography on diethylaminoethyl (DEAE)-cellulose and Sephadex 100 columns. The enzyme has a molecular weight of 45 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and of 90 kDa as determined by gel filtration. The purified T. rangeli alpha-mannosidase has a pH optimum ranging between 5 and 6 and a temperature optimum of 37 degrees C. It has activation energy of 8.15 x 10(2) J mol(-1) K(-1). The enzyme has a Michaelis constant (Km) of 99 microM for the 4-methylumbelliferyl-alpha-D-mannopyranoside substrate (MU-alpha-mann). It is strongly inhibited by swainsonine [inhibition constant (Ki) 0.048 microM] and is moderately inhibited by mannose and alpha-D-methylmannopyranoside. The enzyme activity is decreased in the presence of 1 mM Ca2+, Co2+, Cu2+, Fe2+, Fe3+, Hg2+, Mg2+, and Mn2+.

MeSH terms

  • Animals
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Kinetics
  • Mannosidases* / antagonists & inhibitors
  • Mannosidases* / chemistry
  • Mannosidases* / isolation & purification
  • Mannosidases* / metabolism
  • Metals, Heavy / pharmacology
  • Trypanosoma / enzymology*
  • Trypanosoma / growth & development
  • Trypanosomiasis / parasitology
  • alpha-Mannosidase

Substances

  • Metals, Heavy
  • Mannosidases
  • alpha-Mannosidase