Sulfated proteoglycans of rabbit aorta: selective extraction and alternative method for glycosaminoglycan moiety analysis.
Three different solutions containing urea, guanidine hydrochloride, or a detergent mixture were used to extract proteoglycan molecules (PG) metabolically labeled with 35S from normal rabbit aortic tissue. The size distribution of whole sulfated PG and the glycosaminoglycan (GAG) compositions in the three extracts were compared and found to be characteristically determined by the type of solution used for extraction. The spectrum of sulfated PG isolated by each solution was maintained at consecutive extractions of the tissue, even if this was used after another type of solution. The extracts obtained by using the urea- or guanidine-containing solutions contained similar, rather balanced populations of large and small PG, while the detergent-containing buffer was found to preferentially extract smaller, heparan sulfate-rich aortic PG. The selectivity of various extracting solutions could be exploited to obtain preparations enriched in certain types of sulfated PG. On the other hand, one could obtain a larger variety of 35S-labeled PG from the tissue by consecutively using two solutions with different capacities of extraction. The distribution of GAG moieties among PG populations, separated by size chromatography, was investigated by one of the commonly used methods and by a new method. The standard method is based on comparison of the chromatographic profiles of the extract before and after enzymatic digestions, requiring several chromatographic runs for a sample. In the alternative method proposed, the fractions obtained after a single chromatographic separation are adsorbed onto a support membrane. Processing of the whole membrane by GAG-specific, enzymatic treatments allows simultaneous assessment of GAG types in each fraction.(ABSTRACT TRUNCATED AT 250 WORDS)
Galis
Z S
ZS
Department of Pathology, McGill University, Montreal, Canada.
Alavi
M Z
MZ
Moore
S
S
D000818:Animals; D001011:Aorta / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000032:analysis*; D008970:Molecular Weight; D011509:Proteoglycans / Q000032:analysis*; D011817:Rabbits; D013431:Sulfates / Q000032:analysis; D014020:Tissue Extracts
D006025:Glycosaminoglycans; D011509:Proteoglycans; D013431:Sulfates; D014020:Tissue Extracts