In vivo turnover of the basement membrane and other heparan sulfate proteoglycans of rat glomerulus.

L A Beavan; M Davies; J R Couchman; M A Williams; R M Mason

In vivo turnover of the basement membrane and other heparan sulfate proteoglycans of rat glomerulus.

L A Beavan, M Davies, J R Couchman, M A Williams, R M Mason

Nutritional Immunology

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

48 Citationer (Scopus)

Abstract

Udgivelsesdato: 1989-Mar

Originalsprog	Engelsk
Tidsskrift	Archives of Biochemistry and Biophysics
Vol/bind	269
Udgave nummer	2
Sider (fra-til)	576-85
Antal sider	9
ISSN	0003-9861
Status	Udgivet - 1989

Bibliografisk note

Keywords: Animals; Autoradiography; Basement Membrane; Female; Glycoproteins; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Kidney Glomerulus; Kinetics; Proteochondroitin Sulfates; Proteoglycans; Rats; Rats, Inbred Strains; Sulfur Radioisotopes

Citationsformater

@article{bfa36540598211dd8d9f000ea68e967b,

title = "In vivo turnover of the basement membrane and other heparan sulfate proteoglycans of rat glomerulus.",

abstract = "The metabolic turnover of rat glomerular proteoglycans in vivo was investigated. Newly synthesized proteoglycans were labeled during a 7-h period after injecting sodium [35S]sulfate intraperitoneally. At the end of the labeling period a chase dose of sodium sulfate was given. Subsequently at defined times (0-163 h) the kidneys were perfused in situ with 0.01% cetylpyridinium chloride in phosphate-buffered saline to maximize the recovery of 35S-proteoglycans. Glomeruli were isolated from the renal cortex and analyzed for 35S-proteoglycans by autoradiographic, biochemical, and immunochemical methods. Grain counting of autoradiographs revealed a complex turnover pattern of 35S-labeled macromolecules, commencing with a rapid phase followed by a slower phase. Biochemical analysis confirmed the biphasic pattern and showed that the total population of [35S]heparan sulfate proteoglycans had a metabolic half-life (t1/2) of 20 and 60 h in the early and late phases, respectively. Heparan sulfate proteoglycans accounted for 80% of total 35S-proteoglycans, the remainder being chondroitin/dermatan sulfate proteoglycans. Whole glomeruli were extracted with 4% 3-[(cholamidopropyl)dimethy-lammonio]-1-propanesulfonate-4 M guanidine hydrochloride, a procedure which solubilized greater than 95% of the 35S-labeled macromolecules. Of these 11-13% was immunoprecipitated by an antiserum against heparan sulfate proteoglycan which, in immunolocalization experiments, showed specificity for staining the basement membrane of rat glomeruli. Autoradiographic analysis showed that 18% of total radioactivity present at the end of the labeling period was associated with the glomerular basement membrane. The glomerular basement membrane [35S]heparan sulfate proteoglycans, identified by immunoprecipitation, have a very rapid turnover with an initial phase, t1/2 = 5 h, and a later phase t1/2 = 20 h.",

author = "Beavan, {L A} and M Davies and Couchman, {J R} and Williams, {M A} and Mason, {R M}",

note = "Keywords: Animals; Autoradiography; Basement Membrane; Female; Glycoproteins; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Kidney Glomerulus; Kinetics; Proteochondroitin Sulfates; Proteoglycans; Rats; Rats, Inbred Strains; Sulfur Radioisotopes",

year = "1989",

language = "English",

volume = "269",

pages = "576--85",

journal = "Archives of Biochemistry and Biophysics",

issn = "0003-9861",

publisher = "Academic Press",

number = "2",

}

TY - JOUR

T1 - In vivo turnover of the basement membrane and other heparan sulfate proteoglycans of rat glomerulus.

AU - Beavan, L A

AU - Davies, M

AU - Couchman, J R

AU - Williams, M A

AU - Mason, R M

N1 - Keywords: Animals; Autoradiography; Basement Membrane; Female; Glycoproteins; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Kidney Glomerulus; Kinetics; Proteochondroitin Sulfates; Proteoglycans; Rats; Rats, Inbred Strains; Sulfur Radioisotopes

PY - 1989

Y1 - 1989

N2 - The metabolic turnover of rat glomerular proteoglycans in vivo was investigated. Newly synthesized proteoglycans were labeled during a 7-h period after injecting sodium [35S]sulfate intraperitoneally. At the end of the labeling period a chase dose of sodium sulfate was given. Subsequently at defined times (0-163 h) the kidneys were perfused in situ with 0.01% cetylpyridinium chloride in phosphate-buffered saline to maximize the recovery of 35S-proteoglycans. Glomeruli were isolated from the renal cortex and analyzed for 35S-proteoglycans by autoradiographic, biochemical, and immunochemical methods. Grain counting of autoradiographs revealed a complex turnover pattern of 35S-labeled macromolecules, commencing with a rapid phase followed by a slower phase. Biochemical analysis confirmed the biphasic pattern and showed that the total population of [35S]heparan sulfate proteoglycans had a metabolic half-life (t1/2) of 20 and 60 h in the early and late phases, respectively. Heparan sulfate proteoglycans accounted for 80% of total 35S-proteoglycans, the remainder being chondroitin/dermatan sulfate proteoglycans. Whole glomeruli were extracted with 4% 3-[(cholamidopropyl)dimethy-lammonio]-1-propanesulfonate-4 M guanidine hydrochloride, a procedure which solubilized greater than 95% of the 35S-labeled macromolecules. Of these 11-13% was immunoprecipitated by an antiserum against heparan sulfate proteoglycan which, in immunolocalization experiments, showed specificity for staining the basement membrane of rat glomeruli. Autoradiographic analysis showed that 18% of total radioactivity present at the end of the labeling period was associated with the glomerular basement membrane. The glomerular basement membrane [35S]heparan sulfate proteoglycans, identified by immunoprecipitation, have a very rapid turnover with an initial phase, t1/2 = 5 h, and a later phase t1/2 = 20 h.

AB - The metabolic turnover of rat glomerular proteoglycans in vivo was investigated. Newly synthesized proteoglycans were labeled during a 7-h period after injecting sodium [35S]sulfate intraperitoneally. At the end of the labeling period a chase dose of sodium sulfate was given. Subsequently at defined times (0-163 h) the kidneys were perfused in situ with 0.01% cetylpyridinium chloride in phosphate-buffered saline to maximize the recovery of 35S-proteoglycans. Glomeruli were isolated from the renal cortex and analyzed for 35S-proteoglycans by autoradiographic, biochemical, and immunochemical methods. Grain counting of autoradiographs revealed a complex turnover pattern of 35S-labeled macromolecules, commencing with a rapid phase followed by a slower phase. Biochemical analysis confirmed the biphasic pattern and showed that the total population of [35S]heparan sulfate proteoglycans had a metabolic half-life (t1/2) of 20 and 60 h in the early and late phases, respectively. Heparan sulfate proteoglycans accounted for 80% of total 35S-proteoglycans, the remainder being chondroitin/dermatan sulfate proteoglycans. Whole glomeruli were extracted with 4% 3-[(cholamidopropyl)dimethy-lammonio]-1-propanesulfonate-4 M guanidine hydrochloride, a procedure which solubilized greater than 95% of the 35S-labeled macromolecules. Of these 11-13% was immunoprecipitated by an antiserum against heparan sulfate proteoglycan which, in immunolocalization experiments, showed specificity for staining the basement membrane of rat glomeruli. Autoradiographic analysis showed that 18% of total radioactivity present at the end of the labeling period was associated with the glomerular basement membrane. The glomerular basement membrane [35S]heparan sulfate proteoglycans, identified by immunoprecipitation, have a very rapid turnover with an initial phase, t1/2 = 5 h, and a later phase t1/2 = 20 h.

M3 - Journal article

C2 - 2521995

SN - 0003-9861

VL - 269

SP - 576

EP - 585

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

IS - 2

ER -