Gel Electrophoretic Band Research Articles

Gel electrophoretic analysis of penicillin-binding proteins of coagulase negatibve staphylococci

We analyzed gel electrophoretic banding patterns of penicillin-binding proteins (PBPs) of 16 type strains of coagulase-negative staphylococci (CNS). S. epidermidis, S. haemolyticus, S. saprophyticus, S. hominis, S. xylosus, S. simulans, S. warneri, S. capitis, S. saccharolyticus, S. auricularis, S. caseolyticus, S. gallinarum, S. hycus subsp. hycus, S. cohnii, S. caprae, and S. sciuri subsp. sciuri. The PBP profile of each CNS species was found to be unique and was clearly distinguishable from those of the rest of the species. Together with the previous work of other researchers, this study substantiates the applicability of the PBP profile analysis to the identification of clinical CNS strains.

The presence and origin of phosphopeptides in human saliva.

The presence of phosphopeptides in whole saliva (saliva expectorated from the mouth) was demonstrated and their origin was evaluated. Whole saliva contained much larger numbers of small phosphopeptides than are found in the glandular secretions. Most of these originated from the acidic proline-rich proteins (PRPs) in the major salivary glands and were formed, after secretion into the oral cavity, as a result of rapid degradation by proteolytic enzymes from extraglandular sources contained in sediment from whole saliva. Some peptides may have been formed by cleavage of basic PRPs, but other phosphoproteins apparently contributed little to the observed phosphopeptides. Most of the enzymes that produced phosphopeptides are serine proteinases. The gel-electrophoretic band patterns of the phosphopeptides obtained from 26 individuals of various acidic-PRP phenotypes were remarkably similar, demonstrating that the enzymes responsible were generally present in the population surveyed and that similar cleavages occur regardless of the nature of the acidic PRPs. Many of these peptides were N-terminal proteolytic cleavage products of acidic PRPs. The N-terminal phosphorylated region of acidic PRPs contains various biological activities, such as inhibition of hydroxyapatite formation, calcium binding and binding to hydroxyapatite, the major mineral of teeth. The demonstration of these phosphopeptides in the saliva that is in contact with the oral surface may therefore be of biological importance.

Haemolymph of femaleOxya hyla hyla serville (Orttioptera: Acrididae): A preliminary study

The preliminary light microscopic and biochemical study of the haemolymph ofOxya hyla hyla records the presence of five different types of haemocytes and gives a quantiative estimation of plasma proteins along with their gel electrophoretic band patterns. The higher number of total haemocyte count in adult female compared to that of immature one may be attributed to some kind of synthesis and transport of proteins or other yolk materials during vitellogenesis. High plasma protein concentration during monsoon may be associated with the reproductive maturity of these insects. The gel electrophoretic pattern of plasma proteins reveals the constant presence of two particular bands suggesting the occurrence of lipoprotein and juvenile hormone carrier protein.

Rapid nucleic acid sequencing methodsalternative approaches to facilitating learning

As part of an advanced course on recombinant DNA technology, our students are required to understand the underlying principles relating to a variety of rapid sequencing techniques for both DNA and RNA. To assess the effectiveness of the learning, the students are requested to (i) produce written accounts of the experimental protocols for each method and (ii) demonstrate their understanding of each technique by correctly interpreting schematic gel electrophoretic band patterns. These band patterns are generated by computer and can be displayed in a variety of presentation formats including print, print plus scrambled print, interactive computer-based instruction, and high resolution computer graphics. The results of such an approach have been very encouraging indeed, both from the point of view of student motivation and examination performance and student/staff interest level

Myosin from Striated Adductor Muscle of Chlamys nipponensis akazara

Myosin was isolated from striated adductor muscle of Akazara shell-fish, and purified on DEAE-Sephadex A50. The sedimentation constant (s 20,2 0 W) and the intrinsic viscosity, [eta] of Akazara myosin thus purified were estimated to be 6.6 S and 2.10 dl/g, respectively. In many respects, Akazara myosin was similar to scallop myosin. (1) Only one size of light-chain component (17,000 daltons) was detectable in SDS-gel electrophoresis of Akazara myosin, but two types of light-chain component were seen in urea-gel electrophoresis; these were equivalent to EDTA-light chain and SH-light chain of scallop myosin. The molar ratio of heavy chain (206,000 daltons), EDTA-light chain, and SH-light chain in Akazara myosin was estimated, from the staining densities of gel-electrophoretic bands, to be approximately 1 : 1 : 1. (2) EDTA-washing procedure removed EDTA-light chain only, causing desensitization of Akazara myosin. EDTA-light chain isolated from Akazara myofibrils was able to resensitize EDTA-washed Akazara myosin. Akazara myosin, however, was found to be different from scallop myosin in two important properties: (1) complete removal of EDTA-light chains was required to achieve a complete loss of calcium sensitivity, and full resensitization was attained on recombination of EDTA-light chains with desensitized myosin prepared essentially free from EDTA-light chains. (2) EDTA-light chains isolated from Akazara myofibrils show a calcium-induced UV absorption difference spectrum.

The interaction of polymeric viologens with hydrogenases from Desulfovibrio desulfuricans and Clostridium pasteurianum

The interaction between hydrogenases from either Desulfovibrio desulfuricans or Clostridium pasteurianum and electron donors methyl viologen or polymeric viologens was examined. Extracts from each organism contained a single gel electophoretic band of active hydrogenase. The hydrogenase of D. desulfuricans was much more stable than that of Cl. pasteurianum. With methyl viologen apparent Km and Vm values were 0.5 mM and 0.62 mumole H2/min per milligram protein for the Cl. pasteurianum and 0.7 and 6.2 mumole H2/min per milligram protein, respectively, for the D. desulfuricans enzyme. The hydrogenases bound the polymeric viologens more tightly than methyl viologen, more so for the enzyme of D. desulfuricans than for Cl. pasteurianum. Maximal rate of hydrogen production was less with the polymeric than with methyl viologen. The results suggest that the D. desulfuricans enzyme in conjunction wiion than that from Cl. pasteurianum.

The induction, purification and characterization of 17 beta-hydroxy-C19-steroid dehydrogenase of the female guinea pig kidney.

The female guinea pig kidney cytosol and microsomes contain a trace or no detectable NADP+ or NAD+-linked dehydrogenase activity for several hydroxy-C19-steroids. The administration of testosterone induced a gradual increase in the 17β-C19-steroid dehydrogenase activity of the cytosol, but not of the microsomes, to the level in the male after 50 days. The induced enzyme was purified by (NH4)2SO4 precipitation, Sephadex G-75 nitration and DEAE-Sephadex A-50 and CM-Sephadex C-50 chromatography to give a protein which gave a molecular weight of 32,000 and a single protein and enzyme staining band on gel electrophoresis. The pure enzyme dissociated into three gel electrophoretic bands on removal of 2-mercaptoethanol from the solution and was restored by replacement of the mercaptoethanol. The enzyme in the cytosol, however, dissociated on storage at 4°C for 48 h or longer in the presence or absence of 2-mercaptoethanol. The KM value, steroid specificity, gel electrophoretic pattern, isoelectric value, pH optimum and NADP+ and NAD+ requirement were practically identical with those of the purified major 17β-hydroxy-C19-steroid dehydrogenase of the male guinea pig kidney. The several hydroxy-C19-steroid dehydrogenase activities of the liver were not influenced by testosterone administration. The highest enzyme activity was exhibited with 5β-dihydrotestosterone as substrate.

The Influence of Growing Temperature on the Amino Acid and Nucleic Acid Concentrations of Potato Tubers1

Abstract Seven experimental selections and one commercial cultivar of potato (Solanum tuberosum L.) were grown in moderate (22/18°C, day/night) and cool (11/7°, day/night) regimes to determine the influence of temperature on potato tuber nitrogen, amino acid, and nucleic acid levels. Tubers from low temperatures had higher percentages of rRNA and lower levels of alpha-keto acids. Moderate and cool grown tubers of the same genotype exhibited slightly different gel electrophoretic banding patterns in 88% of the samples, evidence that tuber protein composition was altered. Glutamate, from HCl hydrolysates, was higher in cool grown tubers. Most cool grown tubers were higher in Kjeldahl nitrogen, due in part to higher levels of glutamate family amino acids, i.e. glutamate, proline and arginine. The free amino acids, histidine and arginine were higher in the cool grown tubers. The essential amino acid scores of leucine, isoleucine, lysine, threonine and valine and the amino acid patterns of methionine were significantly lower in the cool grown tubers; consequently, the cool grown tubers were nutritionally inferior per unit of tuber nitrogen.

Isolation and Characterization of Plasminogen Activator from Pig Leucocytes

SummaryPlasminogen activator was isolated from disrupted pig leucocytes by the aid of DEAE chromatography, gel filtration on Sephadex G-100 and final purification on CM cellulose, or by preparative gel electrophoresis.Isolated plasminogen activator corresponds No. 3 band of the starting sample of leucocyte cells (that is composed from 10 gel electrophoretic bands).pH optimum was found to be in pH range 8.0–8.5 and the highest pH stability is between pH range 5.0–8.0.Inhibition studies of isolated plasminogen activator were performed with EACA, AMCHA, PAMBA and Trasylol, using Anson and Astrup method. By Astrup method 100% inhibition was found with EACA and Trasylol and 30% with AMCHA. PAMBA gave 60% inhibition already at concentration 10–3 M/ml. Molecular weight of plasminogen activator was determined by gel filtration on Sephadex G-100. The value obtained from 4 different samples was found to be 28000–30500.

Changing patterns of nucleic acids, basic and acidic proteins in generative and vegetative nuclei during pollen germination and pollen tube growth in Hippeastrum belladonna

Abstract Generative and vegetative nuclei of mature and germinated pollen grains from Hippeastrum belladonna were separated in a continuous Ficoll gradient. Less than 3% contamination was observed between the generative and vegetative nuclear fractions. The vegetative nuclei were composed of two populations; the larger population consisted of nuclei with 1C levels of DNA and the smaller with 2C levels. The generative nuclei consisted of a homogeneous population composed of nuclei possessing 2C levels of DNA. Histone synthesis did not occur in vegetative nuclei. Changes appeared in the gel-electrophoretic banding patterns of the F1 histones of vegetative nuclei during germination. Changes were not observed in the generative nuclei. A reduction of general proteins and RNA was observed in vegetative nuclei by 20 h of germination. The phenol-soluble nuclear proteins of vegetative nuclei revealed transitions in electrophoretic banding patterns during pollen germination that were greater than those shown by the histones. These changes in the PSNP primarily involved reduced concentrations of certain proteins rather than synthesis of new ones. However, a new band was observed in the electrophoretic pattern of the PSNP of vegetative nuclei after 12 h of pollen tube growth. No transition was seen in the PSNP of generative nuclei during pollen germination and tube growth. The regulatory role of the PSNP in cell differentiation is discussed in the light of these findings.

Disc Electrophoresis of Proteins of Senescing and Fresh Leaves and Petals of Certain Ornamental Plants1

Abstract The acrylamide gel electrophoretic band patterns of proteins from young or fresh and old, senescing leaves or petals of roses, carnations, chrysanthemums, and snapdragons showed generally that with aging the initial and prominent, distinct bands near the origin became less distinct and were located at a distance thus indicating the presence of a more heterogeneous admixture of apparently smaller molecular weight protein units. There was also less protein in the senescing tissues. Kinetin treatment tended to maintain the electrophoretic pattern of leaf proteins similar and comparable to proteins from fresh leaves.

STRUCTURE AND SPECIFICITY OF GUINEA PIG 7S ANTIBODIES.

Additional evidence has been obtained to show that different guinea pig anti-hapten antibodies differ in the structure of their L polypeptide chains. Antibodies from animals immunized with the same hapten conjugated to different carrier proteins gave similar starch gel electrophoretic patterns after dissociation of their chains. In a study of fine differences of specificity, cross-reacting antibodies were found to have some L chains with the same electrophoretic mobility. The multiplicity of L chain bands found in the characteristic starch gel electrophoretic patterns of dissociated anti-DNP antibodies was shown to be a reflection of the heterogeneity of antibodies of slightly different specificities. Reduction and alkylation of the active fragment produced by digestion of antibodies with papain yielded starch gel electrophoretic bands corresponding in mobility to L chains. The results are consistent with the notion that L chains are involved in the acquisition of immunologic specificity.