Selective decreased de novo synthesis of glomerular proteoglycans under the influence of reactive oxygen species

Naoki Kashihara, Yuzo Watanabe, Hirofumi Makino, Elisabeth I. Wallner, Yashpal S. Kanwar

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

The effect of reactive oxygen species on de novo synthesis of heparan sulfate proteoglycans (HSPGs) of the renal glomerulus was investigated in an organ perfusion system. Isolated kidneys were perfused for 7 hr with a medium containing [35S]SuIfate to label sulfated proteoglycans or [35S]methionine to label total glomerular glycoproteins. For the generation of reactive oxygen species, xanthine and xanthine oxidase were included in the perfusion medium, and catalase and superoxide dismutase were used as scavenging agents. Proteoglycans were characterized by Sepharose CL-6B and DEAE-Sephacel chromatographies and SDS/PAGE analysis. The labeled glycoproteins were immunoprecipitated with anti-HSPG, anti-type IV collagen, and anti-laminin, and their specific radioactivities were determined. With exposure to reactive oxygen species, a drastic dose-dependent decrease in de novo synthesis of proteoglycans was seen, and that effect was reversible by catalase treatment. No alterations in the biochemical characteristics of proteoglycans were noted. Immunoprecipitation studies revealed a 16-fold decrease in the synthesis of nascent core peptide of HSPGs, while at comparable concentrations of xanthine and xanthine oxidase, synthesis of type IV collagen and laminin slightly decreased (≈15%). Morphologic studies revealed a 14-fold decrease in [35S]sulfate-associated autoradiographic grains overlying the glomerular basement membrane, a critical component of the ultrafiltration apparatus. Relevance of the selective decreased de novo synthesis of HSPGs of the glomerular basement membrane is discussed in terms of increased glomerular permeability to plasma proteins.

Original languageEnglish
Pages (from-to)6309-6313
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number14
DOIs
Publication statusPublished - Jul 15 1992
Externally publishedYes

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Heparan Sulfate Proteoglycans
Proteoglycans
Reactive Oxygen Species
Glomerular Basement Membrane
Xanthine
Collagen Type IV
Xanthine Oxidase
Laminin
Catalase
Glycoproteins
Perfusion
Kidney
Ultrafiltration
Immunoprecipitation
Methionine
Radioactivity
Sulfates
Superoxide Dismutase
Chromatography
Blood Proteins

Keywords

  • Extracellular matrix

ASJC Scopus subject areas

  • General

Cite this

Selective decreased de novo synthesis of glomerular proteoglycans under the influence of reactive oxygen species. / Kashihara, Naoki; Watanabe, Yuzo; Makino, Hirofumi; Wallner, Elisabeth I.; Kanwar, Yashpal S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 14, 15.07.1992, p. 6309-6313.

Research output: Contribution to journalArticle

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