1965/09/01 by P Andrews · 3 citations
Agricultural and Biological Sciences · Biochemistry, Genetics and Molecular Biology · Chemistry · Medicine · #Biochemistry #Blood properties and coagulation #Chemistry #Chromatography #Enzyme #Gel permeation chromatography #Globular protein #Molecular mass #Organic chemistry #Polymer #Protein purification and stability #Proteins in Food Systems #Sephadex #Size-exclusion chromatography
paper · pdf · doi:10.1042/bj0960595
crossref issued 1965/09/01 · crossref published 1965/09/01 · crossref published-print 1965/09/01 · openalex publication_date 1965/09/01 · crossref created 2015/08/10 · crossref deposited 2021/11/30 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/01 · crossref indexed 2026/08/01
1. Correlation between elution volume, V(e), and molecular weight was investigated for gel filtration of proteins of molecular weights ranging from 3500 (glucagon) to 820000 (alpha-crystallin) on Sephadex G-200 columns at pH7.5. 2. Allowing for uncertainties in the molecular weights, the results for most of the carbohydrate-free globular proteins fitted a smooth V(e)-log(mol.wt.) curve. In the lower part of the molecular-weight range the results were similar to those obtained with Sephadex G-75 and G-100 gels. 3. V(e)-log(mol.wt.) curves based on results with the three gels are taken to represent the behaviour of ;typical' globular proteins, and are proposed as standard data for the uniform interpretation of gel-filtration experiments. 4. Some glycoproteins, including gamma-globulins and fibrinogen, do not conform to the standard relationship. The effect of shape and carbohydrate content on the gel-filtration behaviour of proteins is discussed. 5. As predicted by the theoretical studies of other authors, correlation exists between the gel-filtration behaviour and diffusion coefficients of proteins. 6. The lower molecular-weight limit for complete exclusion of typical globular proteins from Sephadex G-200 varies with the swelling of the gel, but is usually >10(6). 7. The concentration-dependent dissociation of glutamate dehydrogenase was observed in experiments with Sephadex G-200, and the sub-unit molecular weight estimated as 250000. The free sub-units readily lose enzymic activity. 8. Recognition of the atypical gel-filtration behaviour of gamma-globulins necessitates an alteration to several molecular weights previously estimated with Sephadex G-100 (Andrews, 1964). New values are: yeast glucose 6-phosphate dehydrogenase, 128000; bovine intestinal alkaline phosphatase, 130000; Aerobacter aerogenes glycerol dehydrogenase, 140000; milk alkaline phosphatase, 180000.