Purification, properties, partial sequence and evolutionary relationships of marsupial erythrocyte carbonic anhydrase.
Carbonic anhydrase (EC 4.2.1.1) has been purified from the erythrocytes of the tammar wallaby (Macropus eugenii desmarest). The enzyme was separated into four zones of activity. The three major individual forms were isolated as discrete entities. Comparison of substrate specificity, specific activities, kinetic constants and inhibition characteristics indicated that these heteromorphs represented minor post-translational modifications of a single gene product of carbonic anhydrase II type. Double-immunodiffusion and peptide mapping confirmed this proposition. The marsupial enzyme exists as a monomer with a molecular weight of about 29 000 containing one atom of zinc per mole which is much more tightly bound to the enzyme than it is in either human carbonic anhydrase I or II. The wallaby enzyme was, like human carbonic anhydrase II, partially inactivated by p-hydroxymercuribenzoate under conditions not inhibitory for human carbonic anhydrase I. The partial sequence of 51 residues of cyanogen-bromide peptides was sufficiently homologous to allow unambiguous overlap with the sequence of both human carbonic anhydrase I and II isozymes as well as with the recently published sequence of an apparent type I-like enzyme from the turtle. It is clear that the single wallaby erythrocyte carbonic anhydrase belongs to the class of separately evolving type II isozymes which have previously been defined only for placental mammals.
D000086:Acetazolamide / Q000494:pharmacology; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D005075:Biological Evolution; D002256:Carbonic Anhydrases / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification; D002712:Chlorides / Q000494:pharmacology; D004912:Erythrocytes / Q000201:enzymology*; D006358:Hot Temperature; D006801:Humans; D007700:Kinetics; D007614:Macropodidae / Q000097:blood*; D008394:Marsupialia / Q000097:blood*; D010446:Peptide Fragments / Q000032:analysis; D013045:Species Specificity; D014426:Turtles / Q000097:blood
D000596:Amino Acids; D002712:Chlorides; D010446:Peptide Fragments; D002256:Carbonic Anhydrases; D000086:Acetazolamide