PXD013273

PXD013273 is an original dataset announced via ProteomeXchange.

Title	Tandem LysC/trypsin digestion performs efficient in detergent conditions and under constant denaturating environments
Description	All shotgun proteomics experiments rely on efficient proteolysis steps for sensitive peptide/protein identification and quantification. Continuous protocol improvement is therefore a constant topic in proteomics research. Previous reports suggest that the sequential tandem LysC/trypsin digest yields higher recovery of fully tryptic peptides than single-tryptic proteolysis. Based on the previous studies, it is assumed that the advantageous effect of tandem proteolysis requires a high sample denaturation state for the initial LysC digest. Therefore, to date, all systematic assessments of LysC/trypsin proteolysis were done in chaotropic environments such as urea. Here we show that the LysC/trypsin digestion can be carried with high efficiency in MS-compatible detergents resulting in higher yields of fully cleaved peptides. We show that higher cleavage efficiency of tandem digests has an impact on absolute protein quantification using iBAQ values. By additionally testing different urea tandem conditions our data implies that beneficial effects of the initial LysC digest do not depend on the sample denaturation state, but more likely, are due to different target specificities of LysC and trypsin. The observed detergent compatibility enables tandem digestion schemes to be implemented in efficient cellular solubilization proteomics procedures without the need for buffer exchange to chaotropic digestion environment.
HostingRepository	PRIDE
AnnounceDate	2020-04-30
AnnouncementXML	Submission_2020-04-30_06:06:45.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD013273
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Timo Glatter
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	Deamidated; Oxidation; Carbamidomethyl
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2019-03-28 00:20:07	ID requested
⏵ 1	2020-04-30 06:06:46	announced

Hakobyan A, Schneider MB, Liesack W, Glatter T, Efficient Tandem LysC/Trypsin Digestion in Detergent Conditions. Proteomics, 19(20):e1900136(2019) [pubmed]

curator keyword: Biological

submitter keyword: LysC, trypsin, protein digestion, shotgun proteomics

Timo Glatter
contact affiliation	Facility for Proteomics and Mass Spectrometry Max-Planck Insitute for Terrestrial Microbiology 35043 Marburg/Germany
contact email	glatter@mpi-marburg.mpg.de
lab head
Timo Glatter
contact affiliation	MPI Marburg
contact email	glatter@mpi-marburg.mpg.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD013273
     1. Label: PRIDE project
     2. Name: Tandem LysC/trypsin digestion performs efficient in detergent conditions and under constant denaturating environments