Glycosaminoglycans from renal brush border membranes of rabbits.

Abstract

Brush border membranes were prepared from the rabbit kidney cortex by sucrose density gradient centrifugation and digested with pronase. A glycosaminoglycan fraction was isolated from the digest by ion-exchange chromatography. Electrophoretogram on cellulose acetate membrane (Separax) of the glycosaminoglycan fraction indicated the presence of two components, which were identified as chondroitin sulfate A/C and dermatan sulfate on the basis of their mobility in Separax electrophoresis and sensitivity to chondroitinase ABC. Neither hyaluronic acid nor heparan sulfate was detected. The results of high-performance liquid chromatography of the pyridylamino derivatives of unsaturated disaccharides in the digests with chondroitinase AC and ABC of the glycosaminoglycan fraction indicated that the ratio of chondroitin 4-sulfate, chondroitin 6-sulfate and dermatan sulfate was 5.5:1.0: 9.1 gycosaminoglycans ; brush border membrane ; kidneyThe brush border membrane is located at cell apex exposed to the lumen of the proximal convoluted tubule, and its function as transportation of various substances in a resorption process has been extensively studied (Aronson and Sacktor 1975).Glycosaminoglycans (GAGS) are polyanionic and hydrophilic substances, and one possible function is to regulate the exchange of water and electrolyte (Murata 1982). GAGS in the kidney have been extensively investigated and it is now established that the mammalian kidney contains heparan sulfate, hyaluronic acid, dermatan sulfate, chondroitin sulfate and oversulfated dermatan sulfate in decreasing amounts in this order, although there may be qualitative and quantita-

Glycosaminoglycans in brush border membranes from rabbit small intestine.

“…The yield was comparable with that from renal brush border membranes (Munakata et al 1985b). The yield of crude glycoconjugates was much lower than that from whole tissues (Munakata et al 1985a).…”

“…As shown in Table 1, the brush border membrane fraction from rabbit small intestine contained all GAGs examined. In contrast, the renal brush border membrane has been shown to lack hyaluronic acid and heparan sulfate, but to contain chondroitin sulfates and dermatan sulfate (Munakata et al 1985b). Although the histological appearance of brush border membranes from the two tissues were similar, their functions are not the same (Critchley et al 1975 ;Aronson and Sacktor 1975).…”

“…In a previous work on glycosaminoglycans (GAGS) from renal brush border membranes of rabbits (Munakata et al 1985b), it was found that the membranes contain chondroitin sulfate and dermatan sulfate, but that hyaluronic acid and heparan sulfate are not detected. Although there have been many informations on GAGS of small intestine, no information is available on GAGS in brush border membranes of this tissue.…”

Changes of glycosaminoglycan composition of uterine myometrium of rabbit induced by female sex steroids.

“…The unsaturated disaccharides released were subjected to reductive amination (Hale et al 1981) and the resulting pyridylamino derivatives were analyzed by HPLC on a column (3.9 x 300 mm) of pBondapak-C18 using Waters Liquid Chromatograph as described previously (Kodama et al 1984). The individual GAG composition was calculated by the method described previously (Munakata et al 1985).…”

“…Each digest was subjected to reductive amination with 2-aminopyridine and The resulting pyridylamino derivatives of unsaturated disaccharides were analyzed by HPLC. The contents of chondroitin 4-and 6-sulfates, and dermatan sulfate were calculated according to the method described previously (Munakata et al 1985) and are summarized in Table 2. Estrogen-treatment increased the contents of chondroitin 4-and 6-sulfates, but decreased that of dermatan sulfate.…”