Age-related changes in carbonic anhydrase of toad lens

Carbonic anhydrase in the toad lens is immunologically identical to the (high activity) enzyme form present in the erythrocyte. As with the erythrocyte carbonic anhydrase, electrofocusing of the freshly-extracted lens enzyme separated three variants with isoelectric points of 6.1, 5.7 and 5.4, all of which exhibited inhibition properties characteristic of the high activity type of carbonic anhydrase. No evidence of the low activity form of the enzyme was found. In contrast to the erythrocyte isoelectric variants, which are stable in vitro, the three variants of lens carbonic anhydrase exhibited progressive anodization in the course of purification and during storage, and could not be isolated in the original form. Acidification of lens carbonic anhydrase appears to be part of the in vivo aging process of the enzyme protein: the still-active enzyme from the oldest lens region--the nucleus--exhibited a considerably lower isoelectric point than the enzyme extracted from the younger, soft fibers of the lens. Although the specific activity of the carbonic anhydrase from the fibers of lens nucleus was considerably lower than the activity of the enzyme from the younger fibers, the observed modification (acidification) of the aged enzyme did not appear to affect substantially its binding properties towards acetazolamide nor the heat lability of the active protein.