Purification and characterization of a lectin from wild sunflower (Helianthus tuberosus L.) tubers.
A lectin (HTTL) was isolated from Helianthus tuberosus L. (wild sunflower) tubers using ion-exchange chromatography, gel filtration, and affinity chromatography. The lectin agglutinated both untreated and trypsin-treated rabbit erythrocytes and did not agglutinate human blood cells of groups A, B, and O. The gel filtration showed the native molecular mass of 72 kDa and subunit molecular masses of 17 and 18.5 kDa on 12% SDS-PAGE. The lectin activity was inhibited by D-mannose. The tetrameric protein revealed a unique characteristic by forming a broad zone of protein in native PAGE at pH 8.3, which dissociated into seven subunits of varying e/m ratios on acid gel at pH 4.3. These seven bands revealed two polypeptide species of molecular masses 17 and 18.5 kDa on 12% SDS-PAGE, as in the case of the native protein. The result indicated that of the seven subunits, three were homotetramers of 17 kDa, one was a homotetramer of 18.5 kDa, and three were heterotetramers of 17 and 18.5 kDa. The lectin was thermostable with broad pH optima (pH 4-8) and had no requirement for divalent metal cations for its activity. The amino acid composition showed that the lectin contained higher amounts of glycine, alanine, and lysine, but no methionine. The sugar content was estimated to be 5.3% mannose equivalent. The HTTL was mitogenic to mouse spleen (total) cells at 25 microg/ml concentration. The lectin showed characteristics different from those of the earlier reported H. tuberosus tuber lectins and hence opens up a new avenue to investigate the structure-function relationship of lectin in Helianthus species.
Suseelan
K N
KN
Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai, India.
Mitra
R
R
Pandey
R
R
Sainis
K B
KB
Krishna
T G
TG
D000372:Agglutination Tests; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000494:pharmacology; D006023:Glycoproteins / Q000494:pharmacology; D006368:Helianthus / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008213:Lymphocyte Activation; D051379:Mice; D008934:Mitogens / Q000494:pharmacology; D037121:Plant Lectins / Q000737:chemistry* / Q000302:isolation & purification* / Q000494:pharmacology; D011817:Rabbits; D013696:Temperature
D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins; D008934:Mitogens; D037121:Plant Lectins