A novel and efficient and low-cost methodology for purification of Macrotyloma axillare (Leguminosae) seed lectin.
The N-acetyl-galactosamine specific lectin from Macrotyloma axillare seeds (LMA) was purified by precipitation and ion exchange chromatography. The LMA 0.2 mol L(-1) fraction showed hemagglutinating activity on erythrocytes A1. The results for molecular mass determinations were about 28 kDa. The LMA pH-dependent assays showed best hemagglutinating activity at pH 6.0-8.0; being decreased at acidic/alkaline conditions and by EDTA treatment. LMA is a tetramer at pH 8.2 and a dimer at pH 4.0. Human erythrocytes from ABO system confirmed the A1 specificity for LMA. This new methodology is useful and easy, with low costs, for lectin purification in large amounts.
de Santana
M A
MA
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, MG, Brazil. marcosefar@yahoo.com.br
Santos
A M C
AM
Oliveira
M E
ME
de Oliveira
J S
JS
Baba
E H
EH
Santoro
M M
MM
de Andrade
M H G
MH
D001671:Biochemistry / Q000191:economics* / Q000379:methods*; D002118:Calcium / Q000494:pharmacology; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D045424:Complex Mixtures / Q000737:chemistry; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol; D007887:Fabaceae / Q000737:chemistry*; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D037121:Plant Lectins / Q000737:chemistry / Q000302:isolation & purification*; D012639:Seeds / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D013696:Temperature
D045424:Complex Mixtures; D037121:Plant Lectins; D000431:Ethanol; D008345:Manganese; D004492:Edetic Acid; D002118:Calcium