Renal Argininosuccinate Synthetase: Purification, Immunohistochemical Localization, and Elastin-Binding Property

Abstract

Several proteins interact with elastin, an essential extracellular matrix element. We describe here an additional elastin-binding protein from the porcine kidney. This protein was extractable with an isotonic buffer, and was purified by a rapid and simple procedure employing its characteristic properties: high affinity for red A-agarose and spontaneous formation of cryoprecipitate. Immunohistochemical studies using antibody against the purified protein demonstrated its localization in the cytoplasm of proximal tubules of the normal rat kidney. On Western blotting using the antibody, an immunoreactive protein was detected in the liver as well as in the kidney. On the basis of the partial amino acid sequences of the purified protein, it was identified as argininosuccinate synthetase. Some biochemical properties of the protein were different from the data reporting a hepatic form of argininosuccinate synthetase, a key enzyme in the urea cycle in the liver. These results suggest that the elastin-binding property of a renal form of the enzyme may be related to a tissue-specific function in the kidney.

Autophagosome-associated variant isoforms of cytosolic enzymes

“…with a size somewhat larger than the 43 kDa autophagosomal isoform (aASS). The (at least) five ASS isoforms previously described from rat liver all have a pI in the alkaline range [37], whereas the pI of aASS (6.7) is clearly acidic, like the kidney isoforms [48]. Because aASS was N-blocked, its Nterminus could not be sequenced by the methods available to us ; whether it is a truncated variant isoform, like aGAPDH and aSCEH, will therefore have to be clarified by future studies.…”

Arginine metabolism: nitric oxide and beyond

“…Approximately 60 % of net arginine synthesis in adult mammals occurs in the kidney [50,60], where citrulline is extracted from the blood and converted stoichiometrically into arginine by the action of ASS and ASL (Figure 2), which are localized within the proximal convoluted tubules [81,[112][113][114][115]. A tight correlation between renal citrulline uptake and renal arginine output has been elegantly demonstrated for both humans and rats [50,116].…”

Arginine and Citrulline and the Immune Response in Sepsis

“…Arginine can be produced from the conversion of citrulline by the combined action of the cytosolic enzymes argininosuccinate synthetase (ASS) and argininosuccinate lyase (ASL) [ 9 , 10 , 20 , 21 , 22 , 23 , 24 ]. Citrulline, the limiting factor in the arginine de novo synthesis [ 9 ], exhibits a low dietary intake (approximately 13% of total arginine) [ 25 ].…”