Abstract
A neutral endopeptidase (EC 3.4.24.5) that degrades alpha- and beta-crystallins occurs in mammalian lens. A procedure for purification of this enzyme from bovine lens is described. The enzyme appears to have a high molecular weight (Mr approximately equal to 700,000) and under denaturing conditions dissociates into at least eight polypeptide subunits with Mrs ranging from 24,000 to 32,000. A neutral proteinase in bovine pituitary has been reported previously to have similar structural characteristics. We have found that this enzyme purified from bovine pituitary is indistinguishable in molecular weight and in subunit composition from bovine lens endopeptidase. In addition, antiserum raised in rabbit against the purified lens enzyme crossreacts with bovine pituitary enzyme. When examined side by side in Ouchterlony double-diffusion tests, the two enzymes give a continuous precipitin line with no spurring. It is concluded that lens neutral endopeptidase and pituitary neutral proteinase are structurally closely similar, if not identical. This is a surprising result because it had been thought previously that the lens endopeptidase was unique to lens, where its crystallin substrates comprise a large proportion of the total tissue protein. In other tissues, crystallin is either absent or occurs, at most, in trace amounts.
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