English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Article Multiplexed proteomics mapping of yeast RNA polymerase II and III allows near-complete sequence coverage and reveals several novel phosphorylation sites.

Mohammed, S., Lorenzen, K., Kerkhoven, R., van Breukelen, B., Vannini, A., Cramer, P., et al. (2008). Article Multiplexed proteomics mapping of yeast RNA polymerase II and III allows near-complete sequence coverage and reveals several novel phosphorylation sites. Analytical Chemistry, 80(10), 3584-3592. doi:10.1021/ac7024283.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0015-7C4F-5 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-2F85-E
Genre: Journal Article

Files

show Files
hide Files
:
1936034.pdf (Publisher version), 3MB
 
File Permalink:
-
Name:
1936034.pdf
Description:
-
Visibility:
Restricted (Max Planck Institute for Biophysical Chemistry (Karl Friedrich Bonhoeffer Institute), Göttingen; )
MIME-Type / Checksum:
application/pdf
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
1936034_Suppl_1.pdf (Supplementary material), 161KB
Name:
1936034_Suppl_1.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
1936034_Suppl_2.pdf (Supplementary material), 25MB
Name:
1936034_Suppl_2.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
1936034_Suppl_3.pdf (Supplementary material), 373KB
Name:
1936034_Suppl_3.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
1936034_Suppl_4.xls (Supplementary material), 119KB
Name:
1936034_Suppl_4.xls
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/vnd.ms-excel / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show
hide
Locator:
http://pubs.acs.org/doi/pdf/10.1021/ac7024283 (Publisher version)
Description:
-

Creators

show
hide
 Creators:
Mohammed, S., Author
Lorenzen, K., Author
Kerkhoven, R., Author
van Breukelen, B., Author
Vannini, A., Author
Cramer, P.1, Author              
Heck, A. J. R., Author
Affiliations:
1Department of Molecular Biology, MPI for Biophysical Chemistry, Max Planck Society, ou_1863498              

Content

show
hide
Free keywords: -
 Abstract: The multisubunit RNA polymerases (Pols) II and III synthesize mainly eukaryotic mRNAs and tRNAs, respectively. Pol II and Pol III are protein complexes consisting of 12 and 17 subunits. Here we analyzed both yeast Pol II and Pol III by multiplexed mass spectrometric analysis using various proteases and both collision induced and electron transfer dissociation. The cumulative data obtained from using the various proteases (trypsin, chymotrypsin, Glu-C and Lys-C) and the two peptide fragmentation approaches allowed us to map nearly the complete sequences of all constituents of both Pol II and III. Notably, chymotrypsin behaved equally well as and in certain circumstances better than trypsin in the context of protein coverage. Although the available high resolution structures have exposed extensive mechanistic insights into transcription, the role of post-translational modification in these processes has been addressed to a lesser extent. In our analysis of Pol II and III we detected 19 phosphorylation sites, of which 12 have not been previously reported. Identified phosphosites were mapped on the Pol II structure which provided indications that they might play a role in regulating the conformation of the clamp region and, as a consequence, interaction of Pol II with nucleic acids. The described multiplexed proteomics approach is generic and reveals that it is possible to map a protein complex to near completion while applying less than 5 μg (approximately 10 pmol) of total starting material.

Details

show
hide
Language(s): eng - English
 Dates: 2008-04-172008
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1021/ac7024283
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Analytical Chemistry
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 80 (10) Sequence Number: - Start / End Page: 3584 - 3592 Identifier: -