Kooperativer Bibliotheksverbund

UID:

Format: XI, 266 S. : Ill., graph. Darst.

ISBN: 9781627033046

Series Statement: Methods in molecular biology 981

Additional Edition: Erscheint auch als Online-Ausgabe 9781627033053

Language: English

URL: Inhaltsverzeichnis

UID:

Format: XI, 266 S. , graph. Darst.

ISBN: 1-62703-304-1 , 978-1-62703-304-6

Series Statement: Methods in molecular biology 981

Note: Includes bibliogaphical references and index

Language: English

Subjects: Biology

URL: Inhaltsverzeichnis

UID:

Format: XI, 266 S. , graph. Darst.

ISBN: 1-62703-304-1 , 978-1-62703-304-6

Series Statement: Methods in molecular biology 981

Note: Includes bibliogaphical references and index

Language: English

Subjects: Biology

URL: Inhaltsverzeichnis

UID:

Format: 1 Online-Ressource (XI, 266 p. 54 illus., 18 illus. in color)

ISBN: 9781627033053

Series Statement: Methods in Molecular Biology, Methods and Protocols 981

Content: Thousands of proteins have been identified to be acetylated. Immense research power has been dedicated to experiments to solve the biological implications of each and every protein acetylation. Two particular sites of protein acetylation have been described intensively: the N-terminal methionine residue of a nascent protein and lysine residues within a protein. In Protein Acetylation: Methods and Protocols, expert researchers in the field detail many of the methods which are now commonly used to study protein acetylation. These include methods and techniques for identification of protein acetylation, column- and gel electrophoresis-based approaches, computationally prediction,  and the biological response to protein acetylation. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.   Authoritative and practical, Protein Acetylation: Methods and Protocols seeks to aid scientists in the further study of the technical aspects involved in understanding protein acetylation

Additional Edition: Erscheint auch als Druck-Ausgabe ISBN 9781627033046

Language: English

DOI: 10.1007/978-1-62703-305-3

URL: Volltext  (URL des Erstveröffentlichers)

UID:

Format: XI, 266 S , Ill., graph. Darst

ISBN: 1627033041 , 9781627033046

Series Statement: Methods in molecular biology 981

Content: Thousands of proteins have been identified to be acetylated. Immense research power has been dedicated to experiments to solve the biological implications of each and every protein acetylation. Two particular sites of protein acetylation have been described intensively: the N-terminal methionine residue of a nascent protein and lysine residues within a protein. In Protein Acetylation: Methods and Protocols, expert researchers in the field detail many of the methods which are now commonly used to study protein acetylation. These include methods and techniques for identification of protein acetylation, column- and gel electrophoresis-based approaches, computationally prediction, and the biological response to protein acetylation. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Protein Acetylation: Methods and Protocols seeks to aid scientists in the further study of the technical aspects involved in understanding protein acetylation. --

Note: Includes bibliogaphical references and index , Validation of protein acetylation by mass spectrometry , Validation of protein acetylation by mass spectrometry , Application of the CIRAD mass spectrometry approach for lysine acetylation site discovery , Application of the MIDAS approach for analysis of lysine acetylation sites , Application of high content biology to yield quantitative spatial proteomic information on protein acetylations , Towards the N-Terminal acetylome : an N-Terminal acetylated peptide enrichment method using CNBr-activated sepharose resin , Identification and analysis of o-acetylated sialoglycoproteins , HPLC-based quantification of in vitro N-terminal acetylation , Separation and purification of multiply acetylated proteins using cation-exchange chromatography , In-gel N-acetylation for the quantification of the degree of protein in vivo N-terminal acetylation , Computational prediction of lysine acetylation proteome-wide , Generation and characterization of pan-specific anti-acetyllysine antibody , Using functional proteome microarrays to study protein lysine acetylation , Quantitation of nucleosome acetylation and other histone posttranslational modifications using microscale NU-ELISA , Preparing semisynthetic and fully synthetic histones H3 and H4 to modify the nucleosome core , Production of amino-terminally acetylated recombinant proteins in E. coli , Identification of lysine acetyltransferase substrates using bioorthogonal chemical proteomics , Nonradioactive in vitro assays for histone deacetylases , Fluorescence-based acetylation assay using thiol-sensitive probes , Analysis of protein acetyltransferase structure-function relation by surface-enhanced raman scattering (SERS): a tool to screen and characterize small molecule modulators , Application of the CIRAD mass spectrometry approach for lysine acetylation site discovery , Application of the MIDAS approach for analysis of lysine acetylation sites , Application of high content biology to yield quantitative spatial proteomic information on protein acetylations , Towards the N-Terminal acetylome : an N-Terminal acetylated peptide enrichment method using CNBr-activated sepharose resin , Identification and analysis of o-acetylated sialoglycoproteins , HPLC-based quantification of in vitro N-terminal acetylation , Separation and purification of multiply acetylated proteins using cation-exchange chromatography , In-gel N-acetylation for the quantification of the degree of protein in vivo N-terminal acetylation , Computational prediction of lysine acetylation proteome-wide , Generation and characterization of pan-specific anti-acetyllysine antibody , Using functional proteome microarrays to study protein lysine acetylation , Quantitation of nucleosome acetylation and other histone posttranslational modifications using microscale NU-ELISA , Preparing semisynthetic and fully synthetic histones H3 and H4 to modify the nucleosome core , Production of amino-terminally acetylated recombinant proteins in E. coli , Identification of lysine acetyltransferase substrates using bioorthogonal chemical proteomics , Nonradioactive in vitro assays for histone deacetylases , Fluorescence-based acetylation assay using thiol-sensitive probes , Analysis of protein acetyltransferase structure-function relation by surface-enhanced raman scattering (SERS): a tool to screen and characterize small molecule modulators

Language: English

UID:

Format: Online-Ressource (XI, 266 p. 54 illus., 18 illus. in color, digital)

ISBN: 9781627033053

Series Statement: Methods in Molecular Biology, Methods and Protocols 981

Language: English

DOI: 10.1007/978-1-62703-305-3

URL: Volltext

URL: Volltext

UID:

Format: XI, 266 Seiten in 1 Teil , 25.4 cm x 17.8 cm, 5308 g

Edition: Softcover reprint of the original 1st edition 2013

ISBN: 9781493959488 , 1493959484

Series Statement: Methods in Molecular Biology 981

Additional Edition: Erscheint auch als Druck-Ausgabe 9781627033046

Language: English

URL: Inhaltstext

URL: http://www.springer.com/

UID:

Format: Online-Ressource (XI, 266 p. 54 illus., 18 illus. in color, digital)

ISBN: 9781627033053

Series Statement: Methods in Molecular Biology, Methods and Protocols 981

Content: Validation of Protein Acetylation by Mass Spectrometry -- Application of the CIRAD Mass Spectrometry Approach for Lysine Acetylation Site Discovery -- Application of the MIDAS Approach for Analysis of Lysine Acetylation Sites -- Application of High Content Biology to Yield Quantitative Spatial Proteomic Information on Protein Acetylations -- Towards the N-terminal Acetylome: An N-terminal Acetylated Peptide Enrichment Method Using CNBr-Activated Sepharose Resin -- Identification and Analysis of O-acetylated Sialoglycoproteins -- HPLC-based Quantification of in vitro N-terminal Acetylation -- Separation and Purification of Multiply Acetylated Proteins using Cation-exchange Chromatography -- In-gel N-acetylation for the Quantification of the Degree of Protein in vivo N-terminal Acetylation -- Computational Prediction of Lysine Acetylation Proteome-wide -- Generation and Characterization of Pan-specific Anti-acetyllysine Antibody -- Using Functional Proteome Microarrays to Study Protein Lysine Acetylation -- Quantitation of Nucleosome Acetylation and other Histone Post-Translational Modifications Using Microscale NU-ELISA -- Preparing Semisynthetic and Fully Synthetic Histones H3 and H4 to Modify the Nucleosome Core -- Production of Amino-terminally Acetylated Recombinant Proteins in E. coli -- Identification of Lysine Acetyltransferase Substrates using Bioorthogonal Chemical Proteomics -- Non-radioactive in-vitro Assays for Histone Deacetylases -- The Fluorescence-Based Acetylation Assay Using Thiol-Sensitive Probes -- Analysis of Protein Acetyltransferase Structure-function Relation by Surface-Enhanced Raman Scattering (SERS): A Tool to Screen and Characterize Small Molecule Modulators.

Content: Thousands of proteins have been identified to be acetylated. Immense research power has been dedicated to experiments to solve the biological implications of each and every protein acetylation. Two particular sites of protein acetylation have been described intensively: the N-terminal methionine residue of a nascent protein and lysine residues within a protein. In Protein Acetylation: Methods and Protocols, expert researchers in the field detail many of the methods which are now commonly used to study protein acetylation. These include methods and techniques for identification of protein acetylation, column- and gel electrophoresis-based approaches, computationally prediction, and the biological response to protein acetylation. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Protein Acetylation: Methods and Protocols seeks to aid scientists in the further study of the technical aspects involved in understanding protein acetylation.

Additional Edition: 9781627033046

Additional Edition: Buchausg. u.d.T. 978-1-627-03304-6

Language: English

DOI: 10.1007/978-1-62703-305-3

URL: lizenzpflichtig

URL: Cover

UID:

Format: Online-Ressource

Content: Fluorescent fusion proteins are widely used to study protein localization and interaction dynamics in living cells. However, to fully characterize proteins and to understand their function it is crucial to determine biochemical characteristics such as enzymatic activity and binding specificity. Here we demonstrate an easy, reliable and versatile medium/high-throughput method to study biochemical and functional characteristics of fluorescent fusion proteins. Using a new system based on 96-well micro plates comprising an immobilized GFP-binding protein (GFP-mulitTrap), we performed fast and efficient one-step purification of different GFP- and YFP-fusion proteins from crude cell lysate. After immobilization we determined highly reproducible binding ratios of cellular expressed GFP-fusion proteins to histone-tail peptides, DNA or selected RFP-fusion proteins. In particular, we found Cbx1 preferentially binding to di-and trimethylated H3K9 that is abolished by phosphorylation of the adjacent serine. DNA binding assays showed, that the MBD domain of MeCP2 discriminates between fully methylated over unmethylated DNA and protein-protein interactions studies demonstrate, that the PBD domain of Dnmt1 is essential for binding to PCNA. Moreover, using an ELISA-based approach, we detected endogenous PCNA and histone H3 bound at GFP-fusions. In addition, we quantified the level of H3K4me2 on nucleosomes containing different histone variants. In summary, we present an innovative medium/high-throughput approach to analyse binding specificities of fluroescently labeled fusion proteins and to detect endogenous interacting factors in a fast and reliable manner in vitro.

In: Datenlieferant: Open Access LMU (Ludwig-Maximilians-University Munich)

Language: English

DOI: 10.1371/journal.pone.0036967

URN: urn:nbn:de:bvb:19-epub-15077-7

URL: https://doi.org/10.1371/journal.pone.0036967

URL: https://nbn-resolving.org/urn:nbn:de:bvb:19-epub-15077-7

URL: https://epub.ub.uni-muenchen.de/15077/1/oa_15077.pdf

URL: https://epub.ub.uni-muenchen.de/15077/

UID:

Format: Online-Ressource

ISSN: 1521-4141

Content: Abstract: The class II transactivator (CIITA) regulates expression of the classical and non‐classical MHC class II genes, HLA‐DR, ‐DP, ‐DQ and ‐DM, but not the B cell‐specific HLA‐DO (DO). Here we show that only HLA‐DR expression is completely dependent on CIITA, since residual expression of HLA‐DM, ‐DP and the β chain of DQ was observed in CIITA‐deficient RJ2.2.5 cells. Although DO shows a unique expression pattern compared to other MHC class II genes, prolonged IFN‐γ treatment of HeLa cells induced DOB expression. Similar to all MHC class II promoters, the DOB promoter contains thehighly conserved W, X1, and Y boxes in addition to a putative OCT box. Mutational analysis of the DOB promoter demonstrated that the X1, Y and OCT boxes are necessary for maximum promoter activity.Furthermore, our results demonstrate that CREB‐1, RFXANK and Oct‐2 occupy the DOB promoter in vivo, However, CIITA and Bob‐1 were only minimally recruited. Finally, fusion of Bjab, a DOB‐negative B cell line, with. 174 B cells that lack the complete MHC class II region (including the DO genes), lead to DO expression. These data indicate that the expression of DO is regulated by an unidentifiedfactor in B cells.

In: volume:33

In: number:9

In: year:2003

In: pages:2361-2371

In: extent:11

In: European journal of immunology, Weinheim : Wiley-VCH, 1971-, 33, Heft 9 (2003), 2361-2371 (gesamt 11), 1521-4141

Language: English

DOI: 10.1002/eji.200323795

URN: urn:nbn:de:101:1-2023100407053249255865

URL: https://doi.org/10.1002/eji.200323795

URL: https://nbn-resolving.org/urn:nbn:de:101:1-2023100407053249255865

URL: https://d-nb.info/1304866483/34

URL: https://doi.org/10.1002/eji.200323795