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Determination os sedimentation coefficient distributions for cartilage proteoglycans
Authors:J C Pita  F J Muller  T Oegema  V C Hascall
Institution:1. Veterans Administration Hospital and University of Miami School of Medicine, Department of Medicine, Miami, Florida 33152 USA;2. Departments of Orthopaedics and Biological Chemistry, University of Minnesota School of Medicine, Minneapolis, Minnesota 55455 USA;3. National Institute of Dental Research, Bethesda, Maryland 20034 USA
Abstract:Sedimentation coefficient distributions of widely polydisperse proteoglycan preparations were made using a previously developed transport sedimentation methodology. Boundary stability was improved by centrifuging samples in a preformed CsCl density gradient (0.016 g/cm4). The results were compared with the distributions obtained with an interferometric analytical centrifugation method. When these two techniques were applied to analyze A1 and A1–D1 proteoglycan preparations, results were in substantial agreement with respect to the mean sedimentation coefficients of the peaks, average S value, sedimentation coefficient distribution, skewness, proportion of monomer and aggregates, and linearity of the plot ln(s) versus C extrapolations to zero concentration. The lower solute concentration compatible with the transport (velocity gradient) method makes this technique particularly suitable for studying the details of proteoglycan distribution of molecular sizes, especially for aggregates.
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