Twelve peptides, including eight conservative amino acid residues in the amino acid sequence of hydrophilic S helix of the alternative oxidase (AOX), were synthesized by solid_phase method. The polypeptide was coupled...Twelve peptides, including eight conservative amino acid residues in the amino acid sequence of hydrophilic S helix of the alternative oxidase (AOX), were synthesized by solid_phase method. The polypeptide was coupled to α chymotrypsinogen, and the antibodies against this complex were obtained in rabbit. By using these antibodies, which were raised to immunoreact with total proteins of purified mitochondria from different organs of mung bean (Phaseolous radiatus L.) seedlings, it was found that there were two hybridizable AOX fractions in the mitochondria of mung bean seedlings. Their molecular weight was about 35 kD and 38 kD, respectively. Moreover, among the respiratory parameters obtained in hypocotyl, true leaf and cotyledon of mung bean seedlings true leaf had the highest total respiration ( V t ), alternative pathway (AP) capacity( V alt ) and the activity of AP (ρ V alt ). Hypocotyl V t and ρ V alt were the lowest, but its V t was higher than that of the cotyledon. The activities of total and cyanide_resistant respiration were consistant with the analysis of Western blotting of AOX expression. The highest V t and ρ V alt in true leaf were accompanied by two hybridizable polypeptides of AOX protein. The next was cotyledon V t and ρ V alt with only one 38 kD hybridizable polypeptide of AOX protein. Hypocotyl V t and ρ V alt were the lowest and its immunobloting band was similar to that of the cotyledon, but the expression amount of 38 kD protein was less than that of the cotyledon. The 35 kD AOX may make the main contribution to the true leaf ρ V alt .展开更多
文摘Twelve peptides, including eight conservative amino acid residues in the amino acid sequence of hydrophilic S helix of the alternative oxidase (AOX), were synthesized by solid_phase method. The polypeptide was coupled to α chymotrypsinogen, and the antibodies against this complex were obtained in rabbit. By using these antibodies, which were raised to immunoreact with total proteins of purified mitochondria from different organs of mung bean (Phaseolous radiatus L.) seedlings, it was found that there were two hybridizable AOX fractions in the mitochondria of mung bean seedlings. Their molecular weight was about 35 kD and 38 kD, respectively. Moreover, among the respiratory parameters obtained in hypocotyl, true leaf and cotyledon of mung bean seedlings true leaf had the highest total respiration ( V t ), alternative pathway (AP) capacity( V alt ) and the activity of AP (ρ V alt ). Hypocotyl V t and ρ V alt were the lowest, but its V t was higher than that of the cotyledon. The activities of total and cyanide_resistant respiration were consistant with the analysis of Western blotting of AOX expression. The highest V t and ρ V alt in true leaf were accompanied by two hybridizable polypeptides of AOX protein. The next was cotyledon V t and ρ V alt with only one 38 kD hybridizable polypeptide of AOX protein. Hypocotyl V t and ρ V alt were the lowest and its immunobloting band was similar to that of the cotyledon, but the expression amount of 38 kD protein was less than that of the cotyledon. The 35 kD AOX may make the main contribution to the true leaf ρ V alt .
