Characterization and cellular distribution of acidic peptide and oligosaccharide metal-binding compounds from kidneys.
Two low-molecular-mass Ni-binding fractions first isolated from human kidneys [Templeton & Sarkar (1985) Biochem. J. 230, 35-42] are further characterized. Both components are acidic and are readily separated from each other by gel chromatography on Bio-Gel P-2. After equilibration with 63Ni the largest complex constitutes about 30% of the radioactive 63Ni and is an approx. 3.5 kDa peptide and the smallest species comprise short oligosaccharides containing 70% of the radioactivity. Both of these components are found in human, bovine and porcine kidneys as well as in a porcine proximal tubule-like cell line LLC-PK1. There is a small variation in amino acid composition between species. The oligosaccharides are reducing sugars and contain sulphate, glucosamine, glucuronic acid and iduronic acid with two to four overall negative charges. The monosaccharide composition was determined by h.p.l.c. with pulsed amperometric detection of the acid hydrolysates and by gas chromatography. In the LLC-PK1 cell line the acidic peptide is both intracellular and extracellular, whereas the oligosaccharides are only intracellular. The concentration of extracellular peptide, as measured by 63Ni binding, is found to increase after exposure of the cells to low micromolar concentrations of Ni, whereas the oligosaccharide concentrations, also measured by 63Ni binding, remain constant. The oligosaccharide component is decreased by 40% in the presence of NH4Cl, suggesting that is derived from degradation of internalized heparan sulphate.
Predki
P F
PF
Department of Biochemistry Research, Hospital for Sick Children, Toronto, Ontario, Canada.
Whitfield
D M
DM
Sarkar
B
B
D000368:Aged; D000596:Amino Acids / Q000032:analysis; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003600:Cytosol / Q000737:chemistry; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008670:Metals / Q000378:metabolism*; D009005:Monosaccharides / Q000032:analysis; D009532:Nickel / Q000378:metabolism; D009844:Oligosaccharides / Q000032:analysis / Q000378:metabolism*; D010455:Peptides / Q000032:analysis / Q000378:metabolism*; D013552:Swine
D000596:Amino Acids; D008670:Metals; D009005:Monosaccharides; D009844:Oligosaccharides; D010455:Peptides; D000643:Ammonium Chloride; D009532:Nickel