Kooperativer Bibliotheksverbund

UID:

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

ISBN: 9781627033046

Serie: Methods in molecular biology 981

Weitere Ausg.: Erscheint auch als Online-Ausgabe 9781627033053

Sprache: Englisch

URL: Inhaltsverzeichnis

UID:

Umfang: Getr. Zählung , Ill., graph. Darst.

Anmerkung: München, Univ., Kumulative Habil.-Schr., 2011

Sprache: Deutsch

Schlagwort(e): Hochschulschrift

UID:

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

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

Serie: Methods in molecular biology 981

Anmerkung: Includes bibliogaphical references and index

Sprache: Englisch

Fachgebiete: Biologie

URL: Inhaltsverzeichnis

UID:

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

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

Serie: Methods in molecular biology 981

Anmerkung: Includes bibliogaphical references and index

Sprache: Englisch

Fachgebiete: Biologie

URL: Inhaltsverzeichnis

UID:

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

ISBN: 9781627033053

Serie: Methods in Molecular Biology, Methods and Protocols 981

Inhalt: 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

Weitere Ausg.: Erscheint auch als Druck-Ausgabe ISBN 9781627033046

Sprache: Englisch

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

URL: Volltext  (URL des Erstveröffentlichers)

UID:

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

ISBN: 1627033041 , 9781627033046

Serie: Methods in molecular biology 981

Inhalt: 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. --

Anmerkung: 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

Sprache: Englisch

UID:

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

ISBN: 9781627033053

Serie: Methods in Molecular Biology, Methods and Protocols 981

Sprache: Englisch

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

URL: Volltext

URL: Volltext

UID:

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

Ausgabe: Softcover reprint of the original 1st edition 2013

ISBN: 9781493959488 , 1493959484

Serie: Methods in Molecular Biology 981

Weitere Ausg.: Erscheint auch als Druck-Ausgabe 9781627033046

Sprache: Englisch

URL: Inhaltstext

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

UID:

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

ISBN: 9781627033053

Serie: Methods in Molecular Biology, Methods and Protocols 981

Inhalt: 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.

Inhalt: 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.

Weitere Ausg.: 9781627033046

Weitere Ausg.: Buchausg. u.d.T. 978-1-627-03304-6

Sprache: Englisch

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

URL: lizenzpflichtig

URL: Cover

UID:

Umfang: Online-Ressource

Inhalt: 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)

Sprache: Englisch

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/