The unusual stability of saporin, a candidate for the synthesis of immunotoxins.
Saporin, a monomeric protein extracted from the seeds of Saponaria officinalis, is an enzyme capable of specific depurination of the eukaryotic ribosomes. Because of its toxicity, saporin proved useful for the synthesis of immunotoxins, chimeric conjugates of a toxin and an antibody specifically directed against cancer cells or other targets. In this paper we report a study of the structural properties of saporin in the presence of denaturing agents and/or proteolytic enzymes. We found that saporin is extremely resistant to denaturation by urea or guanidine (up to 4 M), even at relatively high temperature (up to 55 degrees C). Moreover a structural change detected as a reduction of the fluorescence emission of the single Trp residue is reversible and is not paralleled by changes of the far UV CD spectrum, suggesting that even under harsh experimental conditions unfolding is limited. In good agreement with these results, guanidine-treated saporin is not attacked by proteolytic enzymes. The remarkable resistance of saporin to denaturation and proteolysis suggests this protein as an ideal candidate for biotechnological applications.
Santanché
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S
Dipartimento di Scienze Biochimiche Alessandro Rossi Fanelli, Universitá di Roma La Sapienza, Italy.
Bellelli
A
A
Brunori
M
M
D002942:Circular Dichroism; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D019791:Guanidine; D006146:Guanidines; D006863:Hydrogen-Ion Concentration; D000922:Immunotoxins / Q000737:chemistry* / Q000627:therapeutic use; D007700:Kinetics; D009699:N-Glycosyl Hydrolases; D010940:Plant Proteins / Q000737:chemistry* / Q000378:metabolism / Q000627:therapeutic use; D011489:Protein Denaturation; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000627:therapeutic use; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins; D012639:Seeds / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013696:Temperature; D014364:Tryptophan / Q000737:chemistry
D006146:Guanidines; D000922:Immunotoxins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D014364:Tryptophan; D009699:N-Glycosyl Hydrolases; D000076985:Saporins; D010450:Endopeptidases; D019791:Guanidine