Biomaterial Database

Scanning isoelectric focusing in small density-gradient columns. IV. The use of deuterium oxide for preparing the density gradient and its effects on isoelectric points of proteins. 1975

S Fredriksson

The use of deuterium oxide as a substitute for sucrose in preparing density gradients for small isoelectrofocusing columns has been investigated. A density gradient was created directly in a 1.5-ml column by free inter-diffusion of three deuterium oxide solutions for 3 min. The resulting deuterium oxide concentration course (as shown by measurement of the refractive-index gradient) had a very high degree of linearity. Test runs with beta-lactoglobulin and sperm-whale myoglobin showed that the strength and stability of the deuterium oxide density gradient should normally be sufficient for stabilization of protein zones against convection during isoelectric focusing. The isoelectric points of beta-lactoglobulins A and B were found to increase as the concentration ratio of deuterium oxide to water at focusing level increased. Within the limits of experimental error, the pI shift corresponding to a given change in solvent composition was equal for both components. These findings are accounted for in terms of the deuterium isotope effect on the dissociation constants of protolytic groups and the shift in the asymmetry potential of the glass electrode in deuterium oxide as compared with water. The spontaneous reduction of sperm-whale metmyoglobin to ferrous myoglobin, which occurs on prolonged isoelectric focusing, is discussed, as are the benefits and drawbacks of deuterium oxide as a density-gradient solute.

UI	MeSH Term	Description	Entries
D007525	Isoelectric Focusing	Electrophoresis in which a pH gradient is established in a gel medium and proteins migrate until they reach the site (or focus) at which the pH is equal to their isoelectric point.	Electrofocusing,Focusing, Isoelectric
D007782	Lactoglobulins	Globulins of milk obtained from the WHEY.	Lactoglobulin,beta-Lactoglobulin,beta-Lactoglobulin A,beta-Lactoglobulin B,beta-Lactoglobulin C,beta-Lactoglobulin E,beta-Lactoglobulin F,beta-Lactoglobulin G,beta-Lactoglobulin I,beta Lactoglobulin,beta Lactoglobulin A,beta Lactoglobulin B,beta Lactoglobulin C,beta Lactoglobulin E,beta Lactoglobulin F,beta Lactoglobulin G,beta Lactoglobulin I
D009211	Myoglobin	A conjugated protein which is the oxygen-transporting pigment of muscle. It is made up of one globin polypeptide chain and one heme group.
D002417	Cattle	Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.	Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D002499	Centrifugation, Density Gradient	Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)	Centrifugations, Density Gradient,Density Gradient Centrifugation,Density Gradient Centrifugations,Gradient Centrifugation, Density,Gradient Centrifugations, Density
D003903	Deuterium	The stable isotope of hydrogen. It has one neutron and one proton in the nucleus.	Deuterons,Hydrogen-2,Hydrogen 2
D006863	Hydrogen-Ion Concentration	The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH	pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D014907	Whales	Large marine mammals of the order CETACEA. In the past, they were commercially valued for whale oil, for their flesh as human food and in ANIMAL FEED and FERTILIZERS, and for baleen. Today, there is a moratorium on most commercial whaling, as all species are either listed as endangered or threatened.	Beaked Whales,Berardius,Caperea,Dwarf Sperm Whale,Giant Bottle-Nosed Whales,Goose-Beaked Whale,Gray Whale,Mesoplodon,Narwhals,Pygmy Right Whale,Pygmy Sperm Whale,Right Whale, North Atlantic,Right Whale, Southern,Ziphiidae,Ziphius,Eschrichtius robustus,Eubalaena australis,Grey Whale,Monodon monoceros,North Atlantic Right Whale,Beaked Whale,Bottle-Nosed Whale, Giant,Bottle-Nosed Whales, Giant,Dwarf Sperm Whales,Giant Bottle Nosed Whales,Giant Bottle-Nosed Whale,Goose Beaked Whale,Goose-Beaked Whales,Gray Whales,Grey Whales,Narwhal,Pygmy Right Whales,Pygmy Sperm Whales,Right Whale, Pygmy,Right Whales, Pygmy,Right Whales, Southern,Southern Right Whale,Southern Right Whales,Sperm Whale, Dwarf,Sperm Whale, Pygmy,Sperm Whales, Dwarf,Sperm Whales, Pygmy,Whale,Whale, Grey,Whale, Southern Right,Whales, Grey,Whales, Southern Right