English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
 
 
DownloadE-Mail
  Adapting data-independent acquisition for mass spectrometry-based protein site-specific N-glycosylation analysis.

Pan, K. T., Chen, C. C., Urlaub, H., & Khoo, K. H. (2017). Adapting data-independent acquisition for mass spectrometry-based protein site-specific N-glycosylation analysis. Analytical Chemistry, 89(8), 4532-4539. doi:10.1021/acs.analchem.6b04996.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-EA45-2 Version Permalink: http://hdl.handle.net/21.11116/0000-0001-3C04-8
Genre: Journal Article

Files

show Files
hide Files
:
2418538.pdf (Publisher version), 4MB
Name:
2418538.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
2418538_Suppl_1.pdf (Supplementary material), 2MB
Name:
2418538_Suppl_1.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
2418538_Suppl_2.xlsx (Supplementary material), 277KB
Name:
2418538_Suppl_2.xlsx
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/vnd.openxmlformats-officedocument.spreadsheetml.sheet / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
2418538_Suppl_3.xlsx (Supplementary material), 68KB
Name:
2418538_Suppl_3.xlsx
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/vnd.openxmlformats-officedocument.spreadsheetml.sheet / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
2418538_Suppl_4.xlsx (Supplementary material), 22KB
Name:
2418538_Suppl_4.xlsx
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/vnd.openxmlformats-officedocument.spreadsheetml.sheet / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show

Creators

show
hide
 Creators:
Pan, K. T.1, Author              
Chen, C. C., Author
Urlaub, H.1, Author              
Khoo, K. H., Author
Affiliations:
1Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society, ou_578613              

Content

show
hide
Free keywords: -
 Abstract: A hallmark of protein N-glycosylation is extensive heterogeneity associated with each glycosylation site. In human cells, the constituent glycoforms differ mostly in numerous ways of extensions from an invariable trimannosyl core and terminal modifications. The efficient identification of these glycoforms at the glycopeptide level by mass spectrometry (MS) requires a precursor sampling technique that is not dictated by signal intensity or by preset targets during MS2 data acquisition. We show here that the recently developed data-independent acquisition (DIA) approach is best suited to this demanding task. It allows post-acquisition extraction of glycopeptide-specific fragment-ion chromatograms to be aligned with that of precursor MS1 ion by nanoLC elution time. For any target glycoprotein, judicious selection of the most favorable MS1/MS2 transitions can first be determined from prior analysis of a purified surrogate standard that carries similar site-specific glycosylation but may differ in its exact range of glycoforms. Since the MS2 transitions to be used for extracting DIA data is common to that glycosylation site and not dictated by specific MS1 value, our workflow applies equally well to the identification of both targeted and unexpected glycoforms. Using a case example, we show that, in targeted mode, it identified more site-specific glycoforms than the more commonly used data-dependent acquisition method when the amount of the target glycoprotein was limiting in sample of high complexity. In discovery mode, it allows detection, with supporting MS2 evidence, of under-sampled glycoforms and of those that failed to be identified by searching against predefined glycan library owing to unanticipated modifications.

Details

show
hide
Language(s): eng - English
 Dates: 2017-03-292017-04-18
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1021/acs.analchem.6b04996
 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: 89 (8) Sequence Number: - Start / End Page: 4532 - 4539 Identifier: -