Two specimens of human articulage were successively extracted with solutions of phosphate-buffered saline (PBS), 7 M-urea and 4 M-guanidine hydrochloride (Gdn-HCl). Proteoglycans from individual extracts were fractionated by DEAE-Sephacel chromatography and gel chromatography on Sephacryl S-400. The presence of three populations of large proteoglycans was demonstrated in all three extracts by c...

An explant culture of 15 cynomolgus monkey corneas was incubated with [35S]sulfate and [2-3H]mannose as labeling precursors. A 4 M guanidine HCl extract of the corneal stromas was prepared and combined with a 4 M guanidine HCl extract of stromas from 300 unlabeled corneas. The keratan sulfate proteoglycans in the combined extracts were purified by a combination of DEAE-cellulose chromatography,...

Chick limb buds from 4- to 7-day-old embryos and sterna from 14-day-old embryos were labeled with [35S]sulfate, and the sulfated proteoglycans (PGS) were extracted either with associative (0.5 M guanidine . HCl) or dissociative (4.0 M guanidine . HCl) solvents. The existence of several populations of PGS is revealed in each age group when the limb extracts are analyzed on sucrose density gradie...

We have analyzed the nature of RecA protein-RecA protein interactions using an affinity column prepared by coupling RecA protein to an agarose support. When radiolabeled soluble proteins from Escherichia coli are applied to this column, only the labeled RecA protein from the extract was selectively retained and bound tightly to the affinity column. Efficient binding of purified 35S-labeled RecA...

The application of high-resolution analytical techniques to characterize ancient bone proteins requires clean, efficient extraction to obtain high quality data. Here, we evaluated many different protocols from the literature on ostrich cortical bone and moa cortical bone to evaluate their yield and relative purity using the identification of antibody-antigen complexes on enzyme-linked immunosor...

The histidyl-tRNA synthetase from Salmonella typhimurium was purified to homogeneity by methods described previously. The molecular weight of the native enzyme was determined as 78,000 by high speed equilibrium centrifugation and as 92,000 by gel filtration; a subunit molecular weight of about 40,000 was found by centrifugation in 6 M guanidine HCl, and by sodium dodecyl sulfate gel electrophor...

the isolation of the target protein from inclusion bodies (ibs) is a preliminary step to increase protein titer and to maintain its biological activity. in the present study, the effects of various cell lysis methods and the expression temperature was investigated on the improvement of the subsequent purification steps of mecasermin produced in ib. we also investigated the solubilization profil...

Numerous experimental techniques and computational studies, proposed in recent times, have revolutionized the understanding of protein-folding paradigm. The complete understanding of protein folding and intermediates are of medical relevance, as the aggregation of misfolding proteins underlies various diseases, including some neurodegenerative disorders. Here, we describe the unfolding of M-cry...