scholarly journals Non-Collagen Protein in the Dentin Tissue – The Role in the Process of Dentinogenesis

2018 ◽  
Vol 11 (2) ◽  
pp. 843-849
Author(s):  
Dusan Surdilovic ◽  
Prabhu Manickam Natarajan ◽  
Tatjana Ille ◽  
Shisir Ram Shetty
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Critical Role ◽  
Human Tooth ◽  
Spectrometric Analysis ◽  
Large Protein ◽  
Organic Constituent ◽  
Coomassie Blue ◽  
Collagen Protein ◽  
Tooth Dentin ◽  
Dentin Sialoprotein

Collagen forms the major organic constituent of human tooth dentin with non-collagen proteins and proteoglycans contributing for minor fragment. Studies involving the nature of non–collagen proteins are comparatively rarer than the other dentinal components. With this background the authors conducted the present study. To identify the specific and unspecific non-collagen proteins in the dentin. Samples of teeth tissue prepared using guanine hydrochloride in natrium acetate and then subjected to polyacrylamide gel electrophoresis. The samples were visualized using Coomassie Blue coloring technique. The pattern of electrophoresis of dentin fraction indicated large protein component. The mass spectrometric analysis results of the two gel parts confirmed the presence of dentin sialoprotein and bone sialoprotein in the dentin fraction. Difference in the molecular mass was evident between the two protien fractions. The authors suggest that since dentin sialoprotein is specific to dentin and not identified in any other tissue, it may be a unique constituent and may play a critical role in formation of dentin.

The reactivity to N-ethyl maleimide of the subunits of cytochrome oxidase

1977 ◽  
Vol 55 (9) ◽  
pp. 988-994 ◽  
Author(s):  
A. McGeer ◽  
B. Lavers ◽  
G. R. Williams
Keyword(s):  
Electron Transport ◽  
Cytochrome Oxidase ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Blue Staining ◽  
Beef Heart ◽  
Coomassie Blue Staining ◽  
Coomassie Blue

Beef heart cytochrome oxidase (EC 1.9.3.1) prepared in this laboratory consistently presents 10 Coomassie blue staining zones on SDS–polyacrylamide gel electrophoresis. At pH 7.0 only two of these polypeptides (III and VIa) are labelled by radioactive N-ethyl maleimide (NEM). The labelling of VIa is variable and correlates with the activity of particular oxidase preparations. When cytochrome oxidase is isolated from alkylated membranes, either mitochondria or electron transport particles, polypeptide VIa is found not to be labelled; polypeptide III is more strongly labelled than when isolated oxidase is alkylated, and label now appears in polypeptide I which is not alkylated upon treatment of isolated oxidase with NEM.

scholarly journals Proteomics of Culture Filtrate of Prevalent Mycobacterium tuberculosis Strains: 2D-PAGE Map and MALDI-TOF/MS Analysis

2017 ◽  
Vol 22 (9) ◽  
pp. 1142-1149 ◽  
Author(s):  
Gavish Kumar ◽  
Hari Shankar ◽  
Divakar Sharma ◽  
Prashant Sharma ◽  
Deepa Bisht ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Culture Filtrate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Critical Role ◽  
Laboratory Strain ◽  
Proteomic Profiling ◽  
East African ◽  
Mycobacterium Tuberculosis H37rv ◽  
Flight Mass Spectrometry ◽  
Central Asian

Although diverse efforts have been done to identify biomarkers for control of tuberculosis using laboratory strain Mycobacterium tuberculosis H37Rv, the disease still poses a threat to mankind. There are many emerging M. tuberculosis strains, and proteomic profiling of these strains might be important to find out potential targets for diagnosis and/or prevention of tuberculosis. We evaluated the comparative proteomic profiling of culture filtrate (CF) proteins from prevalent M. tuberculosis strains (Central Asian or Delhi type; CAS1_Del, East African-Indian; EAI-3 and Beijing family) by 2D polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization–time-of-flight mass spectrometry. As a result, we could identify 12 CF proteins (Rv0066c, Rv1310, Rv3375, Rv1415, Rv0567, Rv1886c, Rv3803c, Rv3804c, Rv2031c, Rv1038c, Rv2809, and Rv1911c), which were consistently increased in all prevalent M. tuberculosis strains, and interestingly, two CF proteins (Rv2809, Rv1911c) were identified with unknown functions. Consistent increased intensity of these proteins suggests their critical role for survival of prevalent M. tuberculosis isolates, and some of these proteins may also have potential as diagnostic and vaccine candidates for tuberculosis, which needs to be further explored by immunological analysis.

Thrombin-Induced Surface Changes of Human Platelets in Plasma: Their Relation to Serotonin Release

1979 ◽  
Author(s):  
K. Subbarao ◽  
V.V. Kakkar
Keyword(s):  
Membrane Proteins ◽  
Blood Platelets ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Band ◽  
Human Platelets ◽  
Serotonin Release ◽  
Glycoprotein I ◽  
Reversible Aggregation ◽  
Coomassie Blue ◽  
Labeling Pattern

Membrane proteins of both control and thrombin-treated platelets were labeled by NaB3H4, reduction of Schiff bases formed between pyridoxal 5′-phosphate and protein amino groups. Examination of the labeled polypeptides by SDS-polyacrylamide gel electrophoresis and fluorography disclosed a different labeling pattern for thrombin-treated platelets. The distributions of Coomassie blue-stained protein from treated and untreated cells were, by contrast, almost identical. Fluorographs of control platelets showed a single intensely labeled protein band (mol wt 90,000) whereas with cells exposed to thrombin (30-60 milliunits) about 10 protein bands with mol wts ranging from 43,000 to 200,000 were typically present. Among these were: thrombin-sensitive protein (mol wt 188,000), glycoprotein I (mol wt 150,000) and actin (mol wt 43,000). When serotonin release was prevented, either by reversing platelet aggregation with low amounts of ADP (0.1-0.3 μM) or by preincubating with 3',5'-ADP (20 μM), an inhibitor of both ADP- and thrombin-induced platelet function, the labeling patterns on fluorographs were similar to the control. These results indicate that blood platelets can undergo reversible aggregation without major changes in their surface topography, whereas thrombin-induced serotonin release appears related to structural alterations in platelet membrane proteins.

scholarly journals Immunochemical characterization of the platelet-specific alloantigen Leka: a comparative study with the PlA1 alloantigen

Blood ◽  
1984 ◽  
Vol 64 (6) ◽  
pp. 1212-1219 ◽  
Author(s):  
N Kieffer ◽  
B Boizard ◽  
D Didry ◽  
JL Wautier ◽  
AT Nurden
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Human Platelets ◽  
Blue Staining ◽  
Igg Antibody ◽  
Immune Disorder ◽  
Immunochemical Characterization ◽  
Coomassie Blue Staining ◽  
Coomassie Blue

Abstract We report the immunochemical characterization of a new platelet- specific alloantigen detected using an IgG antibody isolated from the serum of a patient with posttransfusion purpura (PTP). In indirect immunoprecipitation experiments, the antibody, termed anti-Leka, predominantly precipitated glycoprotein (GP) IIb from Triton X-100 lysates of normal human platelets. In an immunoblot procedure, which involved the transfer of platelet polypeptides separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to nitrocellulose membrane, anti-Leka bound exclusively to GP IIb. Under identical conditions, four anti-PlA1 antibodies each reacted with GP IIIa. No binding of anti-Leka IgG occurred to Leka (-) platelets or to their separated polypeptides although GP IIb was normally detected by Coomassie blue staining. After electrophoresis of reduced platelet proteins, the Leka determinant was localized to the IIb alpha chain. Thus, unlike the PlA1 antigen, the Leka determinant was not destroyed by disulfide reduction. Analysis of platelets from a patient with Glanzmann's thrombasthenia revealed little or no binding in the GP IIb position. Anti-Leka permitted the identification of 76,000 and 60,000 dalton fragments of GP IIb retained by the platelet following chymotrypsin treatment. Our results further highlight the immunogenicity of the GP IIb-IIIa complex. They also suggest that antibodies against GP IIb can cause the thrombocytopenia observed in PTP and that anti-PlA1 antibodies do not account exclusively for the pathophysiology of this immune disorder.

Characterization of PIG Platelet Granule Proteins

1979 ◽  
Author(s):  
M.H. Fukami ◽  
J.L. Daniel ◽  
J.S. Bauer
Keyword(s):  
Gel Electrophoresis ◽  
Plasma Membranes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Dense Granule ◽  
Release Mechanism ◽  
Molecular Weights ◽  
Dense Granules ◽  
Coomassie Blue ◽  
Pas Staining

Platelet granules contain glycoproteins similar to those found in platelet membranes (Hagen et al , BBA , 445, 21 4 , 1 976 ). Pig platelet granule fractions enriched in mitochondria, α-granules or dense granules were analyzed by SDS Polyacrylamide gel electrophoresis to determine if there are differences among the organelles. In a reduced system (5Ϊ OTT) the proteins of the ï-granules and dense granules showed staining patterns with Coomassie blue that were distinctly different from whole platelets, isolated membranes or mitochondria. In the granules about 10 to 12 bands with less mobility than actin were visualized. Staining with PAS was obtained in bands with apparent molecular weights of 250, 225, 185, 170, 150, 120, 55, 4B and 40 K. The 185 K band appeared to be the same as “thrombin sensitive protein”. The mobility of the 55 and 48 K hands were identical with the B (B) and γ-bands of bovine fibrinogen. The PAS staining of the granule components was more intense than that of whole platelets for the same amount of protein, indicating that granule membranes may be as rich in glycoproteins as external plasma membranes. With both PAS and Coomassie blue, the a-granule and dense granule staining patterns were almost identical. This observation may be relevant to recent studies which showed that both granule types exhibited similar release characteristics, suggesting that they share a common release mechanism. NIH-JSPHS Grant No. 14217

scholarly journals Child murder in the Early Bronze Age: proteomic sex identification of a cold case from Schleinbach, Austria

2020 ◽  
Vol 12 (11) ◽  
Author(s):  
Katharina Rebay-Salisbury ◽  
Lukas Janker ◽  
Doris Pany-Kucera ◽  
Dina Schuster ◽  
Michaela Spannagl-Steiner ◽  
...  
Keyword(s):  
Bronze Age ◽  
Social Relations ◽  
Tooth Enamel ◽  
Sex Identification ◽  
Child Rearing ◽  
Human Tooth ◽  
Early Bronze Age ◽  
Spectrometric Analysis ◽  
Child Murder ◽  
Quantum Leap

Abstract The identification of sex-specific peptides in human tooth enamel by nanoflow liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS) represents a quantum leap for the study of childhood and social relations more generally. Determining sex-related differences in prehistoric child rearing and mortality has been hampered by the insufficient accuracy in determining the biological sex of juveniles. We conducted mass spectrometric analysis to identify sex-specific peptides in the dental enamel of a child from a settlement pit of the Early Bronze Age settlement of Schleinbach, Austria (c. 1950–1850 bc). Four perimortal impression fractures on the skull of a 5–6-year-old child indicate an intentional killing, with a co-buried loom weight as possible murder weapon. Proteomic analysis, conducted for the first time on prehistoric teeth in Austria, determined the child’s sex as male. While we cannot conclusively determine whether the child was the victim of conflicts between village groups or was slain by members of his own community, we suggest that contextual evidence points to the latter. A possible trigger of violence was the follow-on effects of an uncontrolled middle ear infection revealed by an osteological analysis. The boy from Schleinbach highlights the potential for further investigation of gender-biased violence, infanticide and child murder based on the recently developed method of proteomic sex identification.

Polyacrylamide gel electrophoresis of exoenzymes produced by Pseudomonas fluorescens strain W

1971 ◽  
Vol 17 (2) ◽  
pp. 241-248 ◽  
Author(s):  
Harvey Winters ◽  
W. A. Corpe
Keyword(s):  
Pseudomonas Fluorescens ◽  
Bacterial Cell ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Complex Mixture ◽  
Gram Negative ◽  
Coomassie Blue ◽  
The Individual ◽  
Cell Envelopes

Culture filtrates from strain W of Pseudomonas fluorescens that cause lysis of gram-negative bacterial cell envelopes were examined for specific hydrolases. The enzymes were concentrated by ammonium sulfate fractionation and by column fractionation on Sephadex G-100. Attempts to separate the individual hydrolases quantitatively by elution from DEAE-cellulose failed because of the formation of aggregates. Resolution of the individual hydrolases was accomplished by disc gel electrophoresis using polyacrylamide (7.5%) gels buffered at pH 8.9 with Tris-glycine. The enzyme mixture was separated into 13 distinct protein bands which were stained with Coomassie blue. The individual hydrolases were detected either directly on the gels or by assay after elution from gel segments and included four proteinases, three phosphatases, two β-glucosidases, one ribonuclease, one lipase, one esterase, and one catalase. These methods provide a rapid, sensitive technique for the detection of many individual hydrolases in a complex mixture.

Simultaneous Implantation of Dental Implants and Autogenous Human Dentin

2011 ◽  
Vol 493-494 ◽  
pp. 426-429 ◽  
Author(s):  
Junichi Tazaki ◽  
Masaru Murata ◽  
Y. Nakanishi ◽  
M. Ochi ◽  
Y. Hirose ◽  
...  
Keyword(s):  
Dental Implants ◽  
Bone Defect ◽  
Human Tooth ◽  
Bacteriological Examination ◽  
Missing Teeth ◽  
Hno3 Solution ◽  
Human Dentin ◽  
Tooth Dentin ◽  
Missing Tooth ◽  
Previous Clinical Study

In our previous clinical study, autogenous demineralized dentin matrices (DDM) prepared from the functional vital teeth (#38, #41) of thirty-five-year-old female were grafted on the bone defect, using newly developed mill, and then received to the host without troubles. In this study, we implanted the human tooth dentin adjusted previously and the dental implants into the regions of missing tooth simultaneously. Fifty-seven-year-old female presented with missing teeth (#35-#37, #45-#47). First, a non-functional vital tooth (#18) were extracted and cryopreserved immediately. 11 months after extraction, the tooth was crushed by newly developed auto-crash mill using ZrO2 vessel and ZrO2 blade for 1 minute. The crushed granules were demineralized completely in 2% HNO3 solution, rinsed in cold distilled water and lyophilized (granule size: 0.5-2.0mm). The bacteria-free of the DDM were confirmed by the bacteriological examination before use. Drilling of the prospective implant beds were then performed according to the manufacture’s protocol and a screw-type rough surface implants (Nobel Biocare® Mk III) were placed. The adjusted DDM granules were implanted into the bone defect (#45). There are no postoperative complications at 3 years after implantation. This case indicates that the preserved autogenous DDM can be used as collagenous biomaterials with osteoinductive potency.

A Model of Microcrack Development in Human Tooth Dentin Using Data of Microtomography

2020 ◽  
Vol 46 (5) ◽  
pp. 505-509
Author(s):  
T. S. Argunova ◽  
Zh. V. Gudkina ◽  
M. Yu. Gutkin
Keyword(s):  
Human Tooth ◽  
Tooth Dentin ◽  
Using Data

scholarly journals Baculovirus Per Os Infectivity Factor Complex: Components and Assembly

2019 ◽  
Vol 93 (6) ◽  
Author(s):  
Xi Wang ◽  
Yu Shang ◽  
Cheng Chen ◽  
Shurui Liu ◽  
Meng Chang ◽  
...  
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Critical Role ◽  
Previous Knowledge ◽  
Multiprotein Complex ◽  
Core Complex ◽  
Insect Midgut ◽  
Content Type ◽  
The Core ◽  
Liquid Chromatography Tandem Mass ◽  
The Impact

ABSTRACT Baculovirus entry into insect midgut cells is dependent on a multiprotein complex of per os infectivity factors (PIFs) on the envelopes of occlusion-derived virions (ODVs). The structure and assembly of the PIF complex are largely unknown. To reveal the complete members of the complex, a combination of blue native polyacrylamide gel electrophoresis, liquid chromatography-tandem mass spectrometry, and Western blotting was conducted on three different baculoviruses. The results showed that the PIF complex has a molecular mass of ∼500 kDa and consists of nine PIFs, including a newly discovered member (PIF9). To decipher the assembly process, each pif gene was knocked out from the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) genome individually by use of synthetic baculovirus technology, and the impact on PIF complex formation was investigated. Deletion of pif8 resulted in the formation of an ∼400-kDa subcomplex. Deletion of pif0, -4, -6, -7, or -9 resulted in a subcomplex of ∼230 kDa, but deletion of pif1, -2, or -3 abolished formation of any complex. Taken together, our data identified a core complex of ∼230 kDa, consisting of PIF1, -2, and -3. This revised the previous knowledge that the core complex was about 170 kDa and contained PIF1 to -4. Analysis of the PIF complex in cellular fractions suggested that it is assembled in the cytoplasm before being transported to the nucleus and subsequently incorporated into the envelopes of ODVs. Only the full complex, not the subcomplex, is resistant to proteolytic attack, indicating the essentiality of correct complex assembly for oral infection. IMPORTANCE Entry of baculovirus into host insects is mediated by a per os infectivity factor (PIF) complex on the envelopes of occlusion-derived viruses (ODVs). Knowledge of the composition and structure of the PIF complex is fundamental to understanding its mode of action. By using multiple approaches, we determined the complete list of proteins (nine) in the PIF complex. In contrast to previous knowledge in the field, the core complex is revised to ∼230 kDa and consists of PIF1 to -3 but not PIF4. Interestingly, our results suggest that the PIF complex is formed in the cytoplasm prior to its transport to the nucleus and subsequent incorporation into ODVs. Only the full complex is resistant to proteolytic degradation in the insect midgut, implying the critical role of the entire complex. These findings provide the baseline for future studies on the ODV entry mechanism mediated by the multiprotein complex.

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