Abstract
Glutathione S-transferase fromOctopus vulgaris hepatopancreas was purified to apparent homogeneity by single glutathione-Sepharose-4B affinity chromatography with overall yield 46% and purification 249-fold. The enzyme was a homodimer with subunitM r 24,000, which was smaller than that of the octopus lens S-crystallin (M r 27,000) with glutathione-S-transferase-like structure. Both proteins showed substrate specificities similar toα/π-type isozyme of glutathione S-transferase. Under native conditions, both proteins exhibited multiple forms upon polyacrylamide gel electrophoresis or isoelectric focusing, albeit with distinct mobilities; however, only one kind of N-terminal amino acid sequence was determined for the multiple forms of each protein. The hepatopancreatic GST, withpI value 6.6–7.3, dissociated into two monomers in an acidic or alkaline environment. Two amino acid residues, withpK a values 5.69±0.14 and 9.03±0.11 were involved in the subunit interactions of the hepatopancreatic enzyme.
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Abbreviations
- PAGE:
-
polyacrylamide gel electrophoresis
- SDS:
-
sodium dodecyl sulfate
- IEF:
-
isoelectric focusing
- GSH:
-
glutathione
- GST:
-
glutathione S-transferase
- CDNB:
-
1-chloro-2,4-dinitrobenzene
- EA:
-
ethacrynic acid [2,3-dichloro-4-(2-methylenebutyryl) phenoxy)acetic acid]
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Tang, SS., Lin, CC. & Chang, GG. Isolation and characterization of octopus hepatopancreatic glutathione S-transferase. Comparison of digestive gland enzyme with lens S-crystallin. J Protein Chem 13, 609–618 (1994). https://doi.org/10.1007/BF01890459
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DOI: https://doi.org/10.1007/BF01890459