Biochemical and immunochemical characterization of Brassica juncea glyoxalase I.
A homogenous preparation of glyoxalase I (S-lactoylglutathione-lyase, EC 4.4.1.5) was obtained from Brassica juncea seedlings. The enzyme is a heterodimer with 27,000 and 29,000 M(r) subunits and native M(r) of 56,000. The circular dichroic spectra of the protein showed characteristics of a distinctly helical protein, and magnesium affected the secondary structure. It is a zinc metalloenzyme. Amino acid modification studies suggested the involvement of histidine residues in catalysis. Apo-glyoxalase I was reactivated by divalent cations Mn2+ (0.5 Mm) > Mg2+ (5 Mm) > Zn2+ (0.05 Mm) and Ca2+ (0.01 Mm). Monospecific, polyclonal anti-glyoxalase I antibodies were raised, which showed its presence in seeds, roots, hypocotyl, cotyledon and different flower parts. They showed varied degree of cross reactivity with the extracts from various plants, yeast, bacteria and animal system.
Deswal
R
R
School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.
Sopory
S K
SK
D015153:Blotting, Western; D001937:Brassica / Q000201:enzymology*; D002384:Catalysis; D002942:Circular Dichroism; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D007791:Lactoylglutathione Lyase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D010936:Plant Extracts / Q000276:immunology; D017433:Protein Structure, Secondary
D010936:Plant Extracts; D007791:Lactoylglutathione Lyase