The variants of the protein toxins abrin and ricin. A useful guide to understanding the processing events in the toxin transport.
Kinetic data on inhibition of protein synthesis in thymocyte by three abrins and ricin have been obtained. The intrinsic efficiencies of A chains of four toxins to inactivate ribosomes, as analyzed by ki-versus-concentration plots were abrin II, III > ricin > abrin I. The lag times were 90, 66, 75 and 105 min at a 0.0744 nM concentration of each of abrin I, II, III and ricin, respectively. To account for the observed differences in the dose-dependent lag time, functional and structural variables of toxins such as binding efficiency of B chains to receptors and low-pH-induced structural alterations have been analyzed. The association constants obtained by stopped flow studies showed that abrin-I (4.13 x 10(5) M-1 s-1) association with putative receptor (4-methylumbelliferyl-alpha-D-galactoside) is nearly two times more often than abrin III (2.6 x 10(5) M-1 s-1) at 20 degrees C. Equilibrium binding constants of abrin I and II to thymocyte at 37 degrees C were 2.26 x 10(7) M-1 and 2.8 x 107 M-1 respectively. pH-induced structural alterations as studied by a parallel enhancement in 8-anilino-L-naphthalene sulfonate fluorescence revealed a high degree of qualitative similarity. These results taken with a nearly identical concentration-independent lag time (minimum lag of 41-42 min) indicated that the binding efficiencies and internalization efficiencies of these toxins are the same and that the observed difference in the dose-dependent lag time is causally related to the proposed processing event. The rates of reduction of inter-subunit disulfide bond, an obligatory step in the intoxication process, have been measured and compared under a variety of conditions. Intersubunit disulfide reduction of abrin I is fourfold faster than that of abrin II at pH 7.2. The rate of disulfide reduction in abrin I could be decreased 11-fold by adding lactose, compared to that without lactose. The observed differences in the efficiencies of A chains, the dose-dependent lag period, the modulating effect of lactose on the rates of disulfide reduction and similarity in binding properties make the variants a valuable tool to probe the processing events in toxin transport in detail.
Hegde
R
R
Department of Biochemistry, Indian Institute of Science, Bangalore.
Karande
A A
AA
Podder
S K
SK
D000036:Abrin / Q000378:metabolism* / Q000633:toxicity; D000818:Animals; D001692:Biological Transport; D002846:Chromatography, Affinity; D004220:Disulfides / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010084:Oxidation-Reduction; D014176:Protein Biosynthesis; D051381:Rats; D011975:Receptors, Mitogen / Q000378:metabolism; D012276:Ricin / Q000378:metabolism* / Q000633:toxicity; D013050:Spectrometry, Fluorescence; D013601:T-Lymphocytes / Q000378:metabolism*
D004220:Disulfides; D011975:Receptors, Mitogen; C035488:abrin receptor; D000036:Abrin; D012276:Ricin