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    Wyświetlanie 1-6 z 6
    Tytuł:
    Investigation of asparagine deamidation in a SOD1-based biosynthetic human insulin precursor by MALDI-TOF mass spectrometry
    Autorzy:
    Bierczyńska-Krzysik, Anna
    Łopaciuk, Małgorzata
    Pawlak-Morka, Renata
    Stadnik, Dorota
    Powiązania:
    https://bibliotekanauki.pl/articles/1039302.pdf
    Data publikacji:
    2014
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    asparagine deamidation
    insulin precursor
    superoxide dismutase
    peptide mass fingerprinting
    MALDI-TOF MS
    Opis:
    A biosynthetic human insulin precursor displayed enhanced susceptibility to deamidation at one particular site. The present study was undertaken to monitor progress of precursor deamidation at successive manufacturing stages. MALDI-TOF/TOF MS in combination with controlled endoproteinase Glu-C and endoproteinase Asp-N proteolysis was used for rapid and unambiguous determination of deamidated residue within the investigated structure. Close inspection of isotopic distribution patterns of peptides resulting from enzymatic digestion enabled determination of distinct precursor forms occurring during the production process. Asn, Asp, isoAsp and succinimide derivatives of the amino acid at position 26 were unambiguously identified. These modifications are related to the leader peptide of a precursor encompassing amino acid sequence corresponding to that of superoxide dismutase [Cu-Zn] (SOD1 1, EC=1.15.1.1). Monitoring of precursor deamidation process at successive manufacturing stages revealed that the protein folding stage was sufficient for a prominent replacement of asparagine by aspartic and isoaspartic acid and the deamidated human insulin precursor constituted the main manufactured product. Conversion proceeded through a succinimide intermediate. Significant deamidation is associated with the presence of SNG motif and confirms results achieved previously on model peptides. Our findings highlight an essential role of the specific amino acid sequence on accelerated rate of protein deamidation. To our knowledge, this is the first time that such a dramatic change in the relative abundance of Asp and isoAsp resulting from protein deamidation process is reported.
    Źródło:
    Acta Biochimica Polonica; 2014, 61, 2; 349-357
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    A proteomics approach to identify the differential protein level in cardiac muscle of diabetic rat
    Autorzy:
    Karthik, Dhanaraj
    Vijayakumar, Ravichandran
    Pazhanichamy, Kalailingam
    Ravikumar, Sivanesan
    Powiązania:
    https://bibliotekanauki.pl/articles/1039290.pdf
    Data publikacji:
    2014
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    diabetes mellitus
    cardiac muscle proteome
    2D electrophoresis
    MALDI-TOF-MS
    phylogenetic analysis
    Opis:
    Background: Cardiovascular proteomics investigation reveals the characterization and elucidation of the novel therapeutic targets and strategies to prevent the development of heart failure associated diabetic complication by using 2DE and MS. Methods: The experimental animals were made diabetic with a single intraperitoneal injection of alloxan (150 mg/kg of bw). Albino rats were randomly divided into four individual groups: Group-I control (n=6), group-II alloxan-induced diabetic rats, untreated (n=6), group-III (n=6) and group-IV (n=6) alloxan-induced diabetic rats were treated with aqueous and ethanolic extracts of Cynodon dactylon for 15 days, respectively. Animals were euthanized to collect the heart tissues and blood samples. 2DE sample preparation, gel running and staining (n=6: each groups) were performed at the same time to avoid variation. The result of six gel images from each group were analyzed and evaluated as one match set with 2D software (P<0.05). Results: The above experiment revealed two up-regulated proteins in group-II i.e. NTF4 and ETFB. Conclusions: NTF4 is a neuro-protective agent for neuro-degenerative diseases. It will prevent diabetic secondary complications, such as diabetic polyneuropathy and cardiomyopathy. ETFB is active in the mitochondria, the energy-producing centres in cells. It is clear from the experiment that because of up-regulation of ETFB more energy is availabile and the electron transfer for heart during diabetes is possible, what leads to reduce the oxidative stress and free-radical formation. The up-regulated proteins reduced CVD that occurred just before overt hyperglycaemia due to administration of C. dactylon. This approach established the preliminary reference map for decoding cellular mechanisms linked between pathogenesis CVD and diabetes.
    Źródło:
    Acta Biochimica Polonica; 2014, 61, 2; 285-293
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Mass spectrometry based analysis of erythrocyte membrane associated proteins in chronic myeloid leukemia patients in Sri Lanka
    Autorzy:
    Kottahachchi, D. U.
    Ariyaratne, T. R.
    Jayasekera, G. A. U.
    Powiązania:
    https://bibliotekanauki.pl/articles/411827.pdf
    Data publikacji:
    2014
    Wydawca:
    Przedsiębiorstwo Wydawnictw Naukowych Darwin / Scientific Publishing House DARWIN
    Tematy:
    erythrocyte membrane associated proteins
    1-D-SDS-PAGE
    MALDI-TOF MS
    PMF
    hematological diseases
    Opis:
    The research reported in this paper was conducted to analyze erythrocyte membrane associated proteins (ERMBPs) of some of chronic myeloid leukemia (CML) patients and selected individuals of Sri Lanka employing one dimensional sodium dodecyl sulphate poly acrylamide gel electrophoresis (1D-SDS-PAGE) combined with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI–TOF-MS). Erythrocyte membranes from blood were isolated by osmotic lysis, centrifugation and washings. ERMBPs were separated on 1D-SDS-PAGE, visualized by silver staining and the separated protein bands dissected out from the gel and were subjected to digestion by proteolytic enzyme, trypsin, and the resulting peptide mixture was analyzed by MALDI-TOF-MS. Resulting experimental peptide mass values were analyzed by peptide mass fingerprint (PMF) technique. From this analysis 10 ERMBPs including α and β spectrin, ankyrin, band 3, band 4.1, band 4.2, band 7, dematin, actin, 55 KDa erythrocyte membrane protein were identified accurately with their primary structure information. The study was able to provide some evidence for Cathepsin associated cleavage of Band 3 anion transport protein in CML patients reported previously. In addition we were able to detect changes in gel bands between healthy controls and CML patients around the area of 20 kDa in the 1 D-SDS-PAGE. It was identified as nuclear protein Dbf 4 related factor 1 isoform 2. Although erythrocytes are devoid of nuclei, such unexpected nuclear proteins have been identified in previous research. We were successful in identifying several human ERMBPs with available resources. As the identified proteins were known to be related to pathology of some of the hematological diseases, this methodology could be extended to detect the protein changes in erythrocyte membrane protein associated diseases. Therefore, this initial research would at some point lead to discovery of biomarkers to these hematological diseases in Sri Lanka.
    Źródło:
    International Letters of Chemistry, Physics and Astronomy; 2014, 19, 1; 74-86
    2299-3843
    Pojawia się w:
    International Letters of Chemistry, Physics and Astronomy
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Comparative proteomic analysis of Bombyx mori hemolymph and fat body after calorie restriction
    Autorzy:
    Chen, Huiqing
    Li, Yijia
    Chen, Keping
    Yao, Qin
    Li, Guohui
    Wang, Lin
    Powiązania:
    https://bibliotekanauki.pl/articles/1040312.pdf
    Data publikacji:
    2010
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    calorie restriction
    Bombyx mori
    two-dimensional gel electrophoresis
    proteomic analysis
    MALDI-TOF/TOF MS
    Opis:
    Calorie restriction (CR) is known to extend life span from yeast to mammals. To gain an insight into the effects of CR on growth and development of the silkworm Bombyx mori at protein level, we employed comparative proteomic approach to investigate proteomic differences of hemolymph and fat body of the silkworm larvae subjected to CR. Thirty-nine differentially expressed proteins were identified by MALDI TOF/TOF MS. Among them, 19 were from the hemolymph and 20 from the fat body. The hemolymph of the CR group contained two down-regulated and 17 up-regulated proteins, whereas the fat body contained 15 down-regulated and five up-regulated ones. These proteins belonged to those functioning in immune system, in signal transduction and apoptosis, in regulation of growth and development, and in energy metabolism. Our results suggest that CR can alter the expression of proteins related to the above four aspects, implying that these proteins may regulate life span of the silkworm through CR.
    Źródło:
    Acta Biochimica Polonica; 2010, 57, 4; 505-511
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Bacterial species identification
    Autorzy:
    Kshikhundo, Ronald
    Itumhelo, Shayalethu
    Powiązania:
    https://bibliotekanauki.pl/articles/1153736.pdf
    Data publikacji:
    2016
    Wydawca:
    Przedsiębiorstwo Wydawnictw Naukowych Darwin / Scientific Publishing House DARWIN
    Tematy:
    16S rRNA gene
    Bacteria
    Biolog
    Gram staining
    MALDI-TOF MS
    RiboPrinter
    computational tools
    fatty acids
    identification
    metagenomics
    morphology
    Opis:
    The traditional methods of bacterial identification are based on observation of either the morphology of single cells or colony characteristics. However, the adoption of newer and automated methods offers advantage in terms of rapid and reliable identification of bacterial species. The review provides a comprehensive appreciation of new and improved technologies such fatty acid profiling, sequence analysis of the 16S rRNA gene, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), metabolic finger profiling using BIOLOG, ribotyping, together with the computational tools employed for querying the databases that are associated with these identification tools and high throughput genomic sequencing in bacterial identification. It is evident that with the increase in the adoption of new technologies, bacterial identification is becoming easier.
    Źródło:
    World News of Natural Sciences; 2016, 3; 26-38
    2543-5426
    Pojawia się w:
    World News of Natural Sciences
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    The application of MALDI-TOF MS for dermatophyte identification
    Autorzy:
    Dabrowska, I.
    Dworecka-Kaszak, B.
    Powiązania:
    https://bibliotekanauki.pl/articles/6182.pdf
    Data publikacji:
    2014
    Wydawca:
    Polskie Towarzystwo Parazytologiczne
    Tematy:
    application
    MALDI-TOF MS zob.matrix-assisted laser desorption-ionization time-of-flight mass spectrometry
    matrix-assisted laser desorption-ionization time-of-flight mass spectrometry
    dermatophyte
    identification
    detection
    fungi
    mammal
    skin disease
    hair
    nail
    Opis:
    Dermatophytes are keratinolytic fungi responsible for a wide variety of diseases of the skin, nails and hair of mammals. Their identification is often complicated, labor-intensive and time consuming due to the high degree of intra-species morphological similarity, and also requires scientific knowledge and practice. The aim of this study was to demonstrate that MALDI-TOF MS technique may be a faster and more sophisticated method useful for the identification of dermatophytes and mycoses in general.
    Źródło:
    Annals of Parasitology; 2014, 60, 3
    0043-5163
    Pojawia się w:
    Annals of Parasitology
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
      Wyświetlanie 1-6 z 6

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