Elektromorphe phycochromoproteid-Muster: Eine taxonomische identifikationsmethode für Cyanobackterien-Species

The disc-gel electrophoretograms of aqueous crude extracts of organisms containing phycochromoproteins (Cyanobacteria, Rhodo- and Cryptophyta) give colored ‘fingerprints’, which are species specific and in many cases strain specific also (infraspecific variability). It was found that in a slightly acidic system (pH 4,3) the electrophoretograms in one single gel concentration (16% PAA) or the Hedrick-Smith plots for the phycochromoprotein bands represent a chemotaxonomic key for identification of Cyanobacteria species. The results summarized here in a list of chemotaxonomical and numerical data for 23 strains, allow one to distinguish between the size and charge isomers of the investigated biliproteids.     

Characterization of the phosphorylatable myosin light chain in rat uterus

The 20 kDa myosin light chain of 32P-labeled rat uterus exhibited four spots on two-dimensional gel electrophoretograms; the corresponding autoradiograms revealed that three spots were radioactive. Completely dephosphorylated light chain exhibited three spots on electrophoretograms. Serine and threonine residues of the light chain were found to be phosphorylated in the uterus at a ratio of 6 to 1. During contraction, the amount of each phosphoamino acid increased proportionally to the increase in the total phosphate content of the light chain.     

The Precursors Affecting the Composition of Capsaicin and Its Analogues in the Fruits of Capsicum annuum var. annuum cv. Karayatsubusa

The effects of tropomyosin and troponin on the heat-induced gelation of myosin were investigated by SDS-polyacrylamide gel electrophoresis, scanning electron microscopy and gel rigidity assay, in comparisons with natural and desensitized actomyosin. SDS-polyacrylamide gel electrophoretograms revealed that tropomyosin was almost completely removed from each desensitized actomyosin samples while it was retained in natural actomyosin samples. In spite of this, no significant differences were found in rigidity between natural and desensitized actomyosin gels. No differences could be observed in the microstructure of either actomyosin gel. It may, therefore, be concluded that tropomyosin does not affect the gel texture of the actomyosin system.     

Effect of protein degradation on spot M(r) distribution in 2-D gels--a case study of proteolysis during development of Streptomyces coelicolor cultures

Proteolysis, a regulated biological process, is reflected by protein spot molecular weight distribution in 2-D gel electrophoretograms. Here we report studies of Streptomyces cultures as they undergo two different developmental processes involving proteolysis. Systematic changes in protein molecular weight distribution between the control samples and those with high activity of proteases were demonstrated. The observations were supported by a numerical model of degradation and its influence on the M(r) distribution. Simple statistics could be used to distinguish between normal and degradative 2-D gel electrophoretic patterns.     

Assay for protease inhibitors: qualitative measurement and detection in polyacrylamide gel electrophoretograms

A simple and fast method for qualitative measurement of protease inhibitors has been developed. The high sensitivity of the assay is based on a casein-precipitating reaction at pH 5.5. Cellulose paper strips, moistened with protease inhibitor-containing solutions, are applied to the surface of an agar-skim milk gel previously coated with a protease solution. After incubation for 2 h at 37 degrees C, inhibitory activity is indicated by a translucent zone (absence of casein degradation) on a white background of precipitated small casein fragments. In addition to being useful for rapid screening of a large number of complex samples, the method allows the localization of protease inhibitor activity in sodium dodecyl sulfate-polyacrylamide gel electrophoretograms.     

云南高背鲫鱼不同组织同工酶分析

采用聚丙烯酰胺凝胶电泳技术,分析了滇池中云南高背鲫鱼(Carassius auratus)的脑、眼睛、肝脏、心脏、肌肉5种组织中的酯酶(EST)、超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH)3种同工酶的表达模式,并对各种酶的同工酶酶谱表型进行了分析。结果表明,云南高背鲫鱼的同工酶系统具有明显的组织特异性,且同工酶的种类与活性变化与其功能相适应。     

Comparisons of plasma membrane polypeptides from soybean and alfalfa

Polypeptides were solubilized with sodium dodecyl sulfate from plasma membrane vesicles of eight varieties of soybean roots [Glycine max (L.) Merr.] and of cultured alfalfa cells (Medicago sativa L.). The solubilized polypeptides were analysed by 2D-polyacrylamide gel electrophoresis. Apparent isoelectric point and MW values were obtained for 80 soybean plasma membrane polypeptides and 44 alfalfa plasma membrane polypeptides. From these data composite distribution patterns were constructed, which are representative of the soybean or alfalfa 2D-gels, respectively. The results showed that the general polypeptide staining patterns were similar for all the soybean varieties, but some minor differences were evident. The alfalfa electrophoretograms differed markedly from the soybean electrophoretograms in specific details, though some general pattern similarities were noted. The data are discussed in terms of a physiological role for the integral plasma membrane polypeptides and in terms of the potential for distinguishing among soybean varieties and between species at the plasma membrane polypeptide level.     

Presence of type III collagen in guinea-pig dermal scar.

Guinea-pig dermal scar was shown to contain type III collagen, and, from densitometric analysis of gel electrophoretograms, it was shown to have a higher concentration than the surrounding dermis. This finding is consistent with the 'embryonic' nature of newly formed dermal wound tissue, reflected in increased hydroxylation of collagen lysine and the presence of dihydroxylysinonorleucine (after reduction) as the major cross-link.     

Lactobacillus lactis protoplasts at different stages of cell cultivation

Abstract Protein profiles of both protoplasts and cell homogenates of Lactobacillus lactis prepared in the course of cell multiplication were compared. The sodium dodecyl sulfate acrylamide gel electrophoretograms of whole cell homogenates and protoplasts show almost the same complexity. Protoplast protein profiles after 4 hours of growth exhibit new bands in both the low and high molecular mass region of the gel. The changes in protein composition during conversion of the cells into protoplasts are predominantly quantitative in nature.     

Methods for increasing the resolution of two-dimensional protein electrophoresis

A two-dimensional gel elctrophoresis protocol has been developed which provides for a 1.5-to 3-fold increase in the resolution of proteins compared to other frequently used methods. The major variations from previous protocols include increased pore size in the isoelectric focusing gels; cholamidopropyldimethylhydroxypropanesulfonate, a zwitterionic detergent, replaces most of the Nonidet P-40, a nonionic detergent, in the isoelectric focusing gels; no equilibration step is employed between the first and second dimensional separation. The use of a stacking gel in the second dimension has been eliminated; a more efficient and evenly distributed cooling system has been designed for the molecular mass separation, allowing faster migration with higher current. Finally, the crosslinker diacrylylpiperazine is employed which improves protein separation and detection with ammoniacal silver staining. Silver-stained two-dimensional gel electrophoretograms of human plasma and hamster brain tissues and autoradiographs of rat liver cells are compared to the results obtained from previous methods.     

Genetic polymorphism of tear proteins in the rat

Polymorphism of tear proteins was found by agarose gel electrophoresis among inbred strains of rats. The proteins (RTP-1) are inherited as a single autosomoal trait. The locus was designated Rtp-1 (rat tear protein-1) and it had two codominant alleles (Rtp-1a, Rtp-1b). Although we did not find any recombinant between the Rtp-1 and the Mup-1 loci among 67 backcross progeny, we found 3 strains with the recombinant type between them in 33 inbred strains tested. The results suggest that the Rtp-1 locus is very closely linked with the Mup-1 locus, which belongs to rat linkage group II. RTP-1 proteins strongly reacted with anti-MUP-1 A serum on agarose gel electrophoretograms.     

Quantitative measurement of protease activities in slab polyacrylamide gel electrophoretograms

A simple, rapid, and inexpensive method for the quantitative measurement of protease activity in slab polyacrylamide gel electrophoretograms is described. An electrophoretogram containing separated test samples and dilutions of a standard protease was placed in contact with a thin (0.7-mm) agar gel slab containing gelatin substrate. After 30 min incubation at 37°C, the unhydrolyzed gelatin was precipitated with saturated ammonium sulfate for 10 min. The developed substrate slab proteolytic zymogram was placed between two clear plastic sheets for analysis by densitometry at 525 nm. A standard graph of peak height in the densitometric scan against quantity of standard protease in micrograms or in proteolytic units was plotted. When measurements of a test sample were made in six independent experiments from the initial linear part of the standard curves the mean ± standard deviation was 3.2 ± 0.2 proteolytic units.     

Postmortem Accumulation of Tubulin in Postsynaptic Density Preparations

Abstract: Postsynaptic density (PSD) preparations isolated from canine cerebral cortex that had been left at 0–37°C for various times were found to become enriched in two bands in a time- but not temperature-dependent manner. The two bands were identified as tubulin subunits by gel mobility and immunology. Of all the isolated synaptic structures the increase in tubulin occurred primarily in the PSD fraction. The increase of tubulin also occurred in PSD preparations isolated from canine cerebellum and rat forebrain. Results obtained when PSD fractions were isolated from canine brain obtained as rapidly as possible after the death of the animal indicate that the maximum amount of tubulin in the PSD preparations is 2.5% of total Coomassie blue-stained protein as determined by scanning of gel electrophoretograms. These results imply that tubulin is probably not a major structural protein of the PSD as it exists in situ.     

The synthesis and storage of histones during the oogenesis of Xenopus laevis

Further data, including two-dimensional gel electrophoresis and peptide mapping of newly synthesized proteins, confirms the view that oocytes make several types of histone. The newly synthesized histone is present in both nucleus and cytoplasm, but at a higher concentration in the oocyte nucleus and in great excess over the DNA binding sites. The unfertilized egg seems to contain a pool of histones detectable on two-dimensional electrophoretograms. The peptide maps of these proteins are consistent with their identification as histones. The egg contains enough histone to support nuclear replication through most of cleavage.     

Reserve proteins in the developing seeds of xhaynaldoticum sardoum.

In this study the pattern of storage protein deposition in the developing seeds of xHaynaldoticum sardoum was examined. The accumulation of the various protein classes did not differ greatly between the two lines tested (Culmo Pieno and Culmo Vuoto). Nevertheless, some differences in the deposition pattern were revealed by polyacrylamide gel electrophoresis. In particular, the mature seeds of the two lines presented different electrophoretograms for the prolamin and glutelin proteins. These patterns reflect the differences between the two lines, which could play a role in their ability to adapt to environmental changes. This work constitutes part of the Ph. D. thesis submitted by the first author. The study was supported by a grant (40 %) from the Ministero Pubblica Istruzione.     

THE DEGRADATION OF PEPTIDES AND AMINOACYL AMIDES BY GUINEA-PIG BRAIN

Abstract— The ability of guinea-pig brain to hydrolyse peptides and aminoacyl amides was investigated. The majority of hydrolase activity against both peptides and aminoacyl amides tested was found to reside in the 30,000 g supernatant. Starch gel electrophoretograms of the 30,000 g supernatants showed that seven of the twelve peptides tested were each hydrolysed by more than one peptide hydrolase and that each of the peptide hydrolases observed were capable of hydrolysing more than one peptide. The peptide hydrolases in the 30,000 g supernatant fraction of guinea-pig brain were found to have very similar electrophoretic mobilities to the peptide hydrolases in the 30,000 g supernatant fraction from guinea-pig intestinal mucosa. Only one hydrolase with activity against L-Leu-NH, could be detected in 30,000 g supernatant of guinea-pig brain using starch gel electrophoresis followed by staining and its electrophoretic mobility was identical to that of one of the peptide hydrolases in the same fraction.     

Evidence for a methicillin-hydrolysing β-lactamase in Staphylococcus aureus strains with borderline susceptibility to this drug

A new beta-lactamase that hydrolyses methicillin was found in the membrane fraction of two clinical isolates of Staphylococcus aureus with borderline susceptibility to this drug. 'Methicillinase' activity was detected in renatured sodium dodecyl sulfate polyacrylamide gel electrophoretograms of staphylococcal membrane proteins. The enzyme activity appeared to be inducible and was more easily detected using penicillin G (or methicillin) rather than nitrocefin as substrate. Similar activity was not detected in the membrane fraction of a methicillin-susceptible strain. These results suggest that, in the two borderline susceptible strains, rather than a hyperproduction of the penicillinase a specific methicillin-hydrolysing activity is responsible for the borderline susceptible phenotype.     

普通伏翼不同组织3种同工酶的比较

采用聚丙烯酰胺不连续凝胶垂直板活性电泳,分析了普通伏翼的心、肝、肾、胸肌、肺、胃、小肠、舌8种组织的酯酶(EST)、乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)3种同工酶。结果表明:普通伏翼8种组织的3种同工酶存在差异,其分布具有明显的组织特异性,与器官或组织所执行的功能有关。     

Study of the structural polypeptides of Chilo suppressalis iridescent virus (Iridovirus type 6)]

We report a procedure for the purification of Chilo iridescent virus (Iridovirus type 6), an evaluation of the purification procedure, and the results of analyses of the virion proteins by acrylamide gel electrophoresis. Purity was evaluated in three ways, i.e., by analysis of purified virions from artificial mixtures of infected and labeled uninfected larvae, electrophoresis at neutral pH, and electron-microscopic examination. Analysis of the polypeptides of purified CIV gave the following results: (i) after solubilization with SDS-B-mercaptoethanol, 16 polypeptides could be resolved in Coomassie brillant blue-stained electrophoretograms with molecular weights ranging from 18,000 to 115,000; (ii) after solubilization with SDS-urea, 26 polypeptides could be resolved with molecular weights ranging from 10,000 to 230,000 daltons.     

Elsie 4: quantitative computer analysis of sets of two-dimensional gel electrophoretograms

We have developed and refined a system for quantitative computer analysis of two-dimensional polyacrylamide gel electrophoretograms. The system, named Elsie 4, is based on one described by Vo et al. (Anal. Biochem. 112, 258 (1981]. It is highly automated. Elsie 4 can find, and measure the intensity of, almost any spot resolvable on two-dimensional gels, including spots visible only as shoulders off larger spots and spots so close together that there is no "valley" between them. It can automatically match the spot patterns of different gels, potentially without the need for a user to provide landmark matches. The matches between paired gels let us follow the synthesis of any spot through a set of gels. Information about a group of matched spots can be obtained by referring to any spot in the group. There is generally no need for a standard or reference gel. Data for two experiments can be combined and compared by matching any gel in one experiment with any gel in the other. There are ways to automatically find possible mismatches in sets of gels. Scans and the results of the analysis can be shown on an image displayer. The programs use function libraries; this helps ensure consistency and increase portability. The programs and functions can be linked together in many ways; this lets users build custom programs for analysis of specific experiments.