摘要:

AbstractIn contrast to Ferguson plots based on relative mobilities, Ferguson plots of proteins in polyacrylamide gel electrophoresis based on their absolute mobilities were found to be linear under unusual polymerization conditions which yield relatively wide gel fibers and a low total fiber length per unit weight, but not under previously and commonly used conditions. These linear Ferguson plots in gels of 1, 3 and 5% crosslinking intersect at a single gel concentration between 1 and 2 %T (M‐point). It is postulated that the measure of free mobility of the proteins is the M‐point, and not the intercept of their Ferguson plots with the mobility axis as assumed previously. This postulate abolishes the well‐known paradoxical interpretation of the increase with %Cof the linearly extrapolated intercept of the Ferguson plot with the log(mobility) axis (designatedYo) in terms of free mobility. The postulate is also compatible with the interpretation of the points of intersection of the Ferguson plots of oligomeric series of proteins at finite gel concentrations (designated μ‐points) as their common free mo

ISSN:0173-0835    

DOI:10.1002/elps.1150090702

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY

摘要:

AbstractThe previous conditions for the physical characterization of turnip crinkle virus (TCV) by quantitative agarose gel electrophoresis [1, 2] were limiting the method to the microgram load level and were therefore insufficiently sensitive to satisfy the need in many areas of virology for detection of viruses containing single‐stranded RNA at the nanogram level. The present report remedies that defect by presenting a technique compatible with the nanogram load level of such viruses. The technique is based on a reduction of gel thickness and on the use of silver stainin

ISSN:0173-0835    

DOI:10.1002/elps.1150090703

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY

摘要:

AbstractThe45Ca filtration method which allows calcium‐binding proteins to be detected after their adsorption to nitrocellulose filters was used to test conditions for improved Western blotting of calmodulin. Our results indicate that dodecyl sulphate is not efficiently removed from calmodulin during electroelution in standard transfer buffers. This detergent disrupts binding of calmodulin to nitrocellulose. Electrotransfer in potassium buffer which precipitates dodecyl sulphate leads to a 6‐fold increase in calmoculin retention by nitrocellulose. With this and other modifications, the detection limit of the45Ca overlay method of Maruyamaet al. (J. Biochem.1984,95, 511–519) for calmodulin is increased to 0.2 μg pe

ISSN:0173-0835    

DOI:10.1002/elps.1150090704

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY

摘要:

AbstractMolecular mass, Stokes' radius, frictional coefficient and isomer‐type of non‐denatured proteins can be obtained by time‐dependent gradient gel electrophoresis by evaluating the resulting data using a two‐step mathematical procedure. Provided a histochemical staining procedure is available to locate the position of an enzyme in the gel, crude cell extracts can be used for estimating their molecular size properties. The computation of molecular properties of non‐denatured proteins is demonstrated for isozymes of aspartate aminotransferase (EC 2.6.1.1), peroxidase (EC 1.11.1.42) and glucose‐6‐phosphate dehydrogenase (EC 1.1.1.49) from current‐year needles of spruce. The resulting data as well as those which were calculated for esterase (EC 3.1.1.1), glutamate dehydrogenase (EC 1.4.1.4), isocitrate dehydrogenase (EC 1.1.1.42) and shikimate dehydrogenase (EC 1.1.1.25) are in accordance with those reported in the literature. The method described may be applied to various scientific areas such as genetics or environmental pollution. It could be shown here that current‐year needles of injured spruce (damage class 3) contained two more peroxidase isozymes and one more glucose‐6‐phosphate dehydrogenase isozyme than those from non‐injured trees. These differences may mark two genotypes of spruce of different susceptibilities towards present‐da

ISSN:0173-0835    

DOI:10.1002/elps.1150090705

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY

摘要:

AbstractHuman serum β2‐glycoprotein I was analyzed by isoelectric focusing followed by im‐munoprinting or immunoblotting with monospecific antiserum. Isoelectric focusing revealed a heterogeneous pattern consisting of 4 major and 4–5 minor bands with isoelectric points of the major bands between pH 5.4 and 6.2. Comparative analysis of sera from more than 400 healthy blood donors showed individual variations of band patterns: six different phenotypes were observed. A family study of 44 families with a total of 129 children demonstrated the genetic control of this variation. Presumably, three alleles, called B2G*1, B2G*2, and B2G*3, determine six phenotypes: B2G 1, 2, 3, 1–2, 1–3 and 2–3. The phenotype 3, however, has not been found in this study. An additional phenotype, noted in one serum specimen, was tentatively classified as B2G 2–4. The distribution of phenotypes and alleles in two populations, from Munich and from Tyrol, has been examined and the frequencies are presented. This genetic polymorphism appears not to be associated with inherited quantitative variations of β2‐glycoprotein I found earlier. The inherited variations can still be recognized after treatment of sera with neuraminidase and with endoglycosidase F, although the banding pattern is altered and shifted towards the cathode. The genetic polymorphism can, therefore, not be ascribed to variations residing in the carbohy

ISSN:0173-0835    

DOI:10.1002/elps.1150090706

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY

摘要:

AbstractLatex serum proteins fromHevea brasiliensiswere studied by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. Proteins from whole serum and fractions isolated by gel chromatography on Ultrogel AcA 44 were analyzed. No qualitative clonal differences were found in the protein patterns of whole latex or in the fractions but laser densitometry revealed reliable quantitative differences in protein composition. Reproducible mobilities and molecular weights of selected bands were obtained both within single gels as well as in different gels, analyzing severallots of latex received at various times from aHeveaexperimental field station. The clones compared were IAN 710, GV 31, GV 42; the first one had the highest rubber yield

ISSN:0173-0835    

DOI:10.1002/elps.1150090707

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY

摘要:

AbstractFive cellulase components were identified and purified in one step fromStreptomycesstrain A20 using electroendosmotic preparative electrophoresis. By this procedure up to 18 mg of protein mixture could be loaded on the column, with an estimated recovery of 60–70% of total activity; activity and protein recovery could be estimated 32% and 47%, respectively, if only activity peaks were considered. In comparison to other purification methods, this technique results in high protein recovery and resolution of applied sample

ISSN:0173-0835    

DOI:10.1002/elps.1150090708

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY

摘要:

AbstractA new method for visualising proteins in two‐dimensional polyacrylamide gels was developed. Proteins were labelled with the fluorophore 2‐methoxy‐2,4‐diphenyl‐3(2H)furanone (MDPF) while present in the first‐dimensional gel after isoelectric focusing and subsequently electrophoresed into the second‐dimensional gel. High resolution spot patterns were produced and compared with other methods of visualisation. A new rapid imaging system based on a cooled charge‐coupled‐device was used to view the two‐dimensional fluorescent protein spot patterns. The method allows the immediate and rapid imaging of two‐dimensional gels at the end of electrophoresis with

ISSN:0173-0835    

DOI:10.1002/elps.1150090709

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY

摘要:

AbstractA method is described which allows to reveal simultaneously the proteolytic patterns of numerous polypeptides separated by two‐dimensional electrophoresis. After two‐dimensional electrophoresis, the gels were dipped successively in buffers for preequilibration, protease digestion, and reequilibration. They were then returned to the electrophoresis tank, and electrophoresis was continued for a short time. After silver staining, digestion products appeared, lined up behind the original polypeptide spots. The method allows proteolytic patterns of numerous polypeptides to be viasualized simply and quickly. Among proteins of wheat leaves, 31 groups of related polypeptides were found according to the similarity of their proteolytic patte

ISSN:0173-0835    

DOI:10.1002/elps.1150090710

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY

摘要:

AbstractHuman skin fibroblasts secrete over 50 proteins into the culture medium. In this paper these are characterised using two‐dimensional polyacrylamide gel electrophoresis and peptide mapping of proteins metabolically labelled in the presence and absence of tunicamycin. Thirty of these proteins have been shown to beN‐glycosides, 4 areO‐glycosides and 10 are not glycosylated. Of the major proteins, groups 1–4 have previously been shown to be fibroblast specific. Peptide mapping and tunicamycin treatment has identified that groups 1 and 2, and 3 and 4 are closely related and that groups 1 and 3 arise byN‐glycosylation of 2 and 4, respectively. The unglycosylated precursor forms of several other proteins have also been identified. This approach to the analysis of protein secretion provides an abundance of information on many proteins simultaneously and can be used to assess the changes in protein secretion associated with development, and to identify extracellular growth factors and other regulatory

ISSN:0173-0835    

DOI:10.1002/elps.1150090711

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1988

数据来源: WILEY