Three distinct molecular species of proteoglycan synthesized by the rat limb bud at the prechondrogenic stage

Journal Article · Wed Nov 15 04:00:00 EST 1989 · Archives of Biochemistry and Biophysics; (USA)

DOI:https://doi.org/10.1016/0003-9861(89)90364-0· OSTI ID:5026821

Matsui, F; Oohira, A; Shoji, R; Nogami, H ^[1]

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1. Central Hospital, Aichi (Japan)

To characterize proteoglycans in the prechondrogenic limb bud, proteoglycans were extracted with 4 M guanidine HCl containing a detergent and protease inhibitors from Day 13 fetal rat limb buds which had been labeled with (35S)sulfate for 3 h in vitro. About 90% of 35S-labeled proteoglycans was solubilized under the conditions used. The proteoglycan preparation was separated by DEAE-Sephacel column chromatography into three peaks; peak I eluted at 0.45 M NaCl concentration, peak II at 0.52 M, and peak III at 1.4 M. Peaks I and III were identified as proteoglycans bearing heparan sulfate side chains. The heparan sulfate proteoglycan in peak III was larger in hydrodynamic size than the proteoglycan in peak I. The heparan sulfate side chains of peak III proteoglycan were smaller in the size and more abundant in N-sulfated glucosamine than those of peak I proteoglycan. Peak II contained a chondroitin sulfate proteoglycan with a core protein of a doublet of Mr 550,000 and 500,000. The chondroitin sulfate proteoglycan was easily solubilized with a physiological salt solution and the heparan sulfate proteoglycan in peak I was partially solubilized with the physiological salt solution. The remainder of the proteoglycan in peak I and the heparan sulfate proteoglycan in peak III could be solubilized effectively only with a solution containing a detergent, such as nonanoyl-N-methylglucamide. This observation indicates the difference in the localization among these three proteoglycans in the developing rat limb bud.

• OSTI ID: 5026821
• Journal Information: Archives of Biochemistry and Biophysics; (USA), Journal Name: Archives of Biochemistry and Biophysics; (USA) Journal Issue: 1 Vol. 275:1; ISSN ABBIA; ISSN 0003-9861
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
BODY AREAS
CARBOXYLIC ACIDS
CHEMICAL COMPOSITION
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ELECTROPHORESIS
EVEN-ODD NUCLEI
GLYCOPROTEINS
ION EXCHANGE CHROMATOGRAPHY
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIMBS
MAMMALS
MOLECULAR WEIGHT
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PROTEINS
RADIOISOTOPES
RATS
RODENTS
SEPARATION PROCESSES
SULFATES
SULFUR 35
SULFUR COMPOUNDS
SULFUR ISOTOPES
TRACER TECHNIQUES
TWO-DIMENSIONAL ELECTROPHORESIS
VERTEBRATES