Partial characterization of proteoglycans synthesized by human glomerular epithelial cells in culture

Journal Article · Wed Feb 28 23:00:00 EST 1990 · Archives of Biochemistry and Biophysics; (USA)

DOI:https://doi.org/10.1016/0003-9861(90)90595-P· OSTI ID:6921455

Klein, D J; Oegema, Jr, T R; Fredeen, T S; van der Woude, F; Kim, Y; Brown, D M ^[1]

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1. Univ. of Minnesota Department of Pediatrics, MN (USA)

Confluent adult and fetal human glomerular epithelial cells were incubated for 24 h in the presence of (3H)-amino acids and (35S)sulfate. Two heparan-35SO4 proteoglycans were released into the culture medium. These 35S-labeled proteoglycans eluted as a single peak from anion exchange chromatographic columns, but were separable by gel filtration on Sepharose CL-6B columns. The larger heparan-35SO4 proteoglycan eluted with the column void volume and at a Kav of 0.26 from Sepharose CL-4B columns. The most abundant medium heparan-35SO4 proteoglycan was a high buoyant density proteoglycan similar in hydrodynamic size (Sepharose CL-6B Kav 0.23) to those previously described in glomerular basement membranes and isolated glomeruli. Heparan-35SO4 chains from both proteoglycans were 36 kDa. A smaller proportion of Sepharose CL-6B excluded dermatan-35SO4 proteoglycan was also synthesized by these cells. The predominant protein cores of both medium heparan-35SO4 proteoglycans were approximately 230 and 180 kDa. A hybrid chondroitin/dermatan-heparan-35SO4 proteoglycan with an 80-kDa protein core copurified with the smaller medium heparan-35SO4 proteoglycan. This 35S-labeled proteoglycan appeared as a diffuse, chondroitinase ABC sensitive 155-kDa fluorographic band in sodium dodecyl sulfate-polyacrylamide gels after the Sepharose CL-6B Kav 0.23 35S-labeled proteoglycan fraction was digested with heparitinase. The heparitinase generated heparan sulfate proteoglycan protein cores and the 155-kDa hybrid proteoglycan fragment had molecular weights similar to those previously identified in rat glomerular basement membrane and glomeruli using antibodies against a basement membrane tumor proteoglycan precursor.

• OSTI ID: 6921455
• Journal Information: Archives of Biochemistry and Biophysics; (USA), Journal Name: Archives of Biochemistry and Biophysics; (USA) Vol. 277:2; ISSN ABBIA; ISSN 0003-9861
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMAL TISSUES
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CELL CULTURES
CHEMICAL COMPOSITION
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ENZYME ACTIVITY
ENZYMES
EPITHELIUM
EVEN-ODD NUCLEI
FETUSES
GLYCOPROTEINS
GLYCOSYL HYDROLASES
HYDROGEN COMPOUNDS
HYDROLASES
HYDROXY ACIDS
ION EXCHANGE CHROMATOGRAPHY
ISOTOPE APPLICATIONS
ISOTOPE DILUTION
ISOTOPES
KIDNEYS
LEUCINE
LIGHT NUCLEI
MAMMALS
MAN
MOLECULAR WEIGHT
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PRIMATES
PROTEINS
RADIOISOTOPES
SEPARATION PROCESSES
SERINE
SULFATES
SULFUR 35
SULFUR COMPOUNDS
SULFUR ISOTOPES
TISSUES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES