PXD002409

PXD002409 is an original dataset announced via ProteomeXchange.

Title	Systematic optimization of C-TAILS approach for deep screening of C-terminomics
Description	It is well known that protein C-termini play important roles in various biological processes, and thus the precise characterization of C-termini is essential for fully elucidating protein structure and understanding protein functions. Although many efforts have been made in the fields during the latest two decades, the progress is still far behind its counterpart, N-termini, and it necessitates more novel or optimized methods. Herein, we reported an optimized C-termini identification approach based on the C-terminal amine-based isotope labeling of substrates (C-TAILS) method. We optimized the amidation reaction conditions to achieve higher yield of fully amidated product. We evaluated different carboxyl and amine blocking reagents and found the superior performance of Ac-NHS and ethanolamine. Replacement of dimethylation with acetylation for Lys blocking resulted in the identification of 232 C-terminal peptides in E. Coli sample, about 42% higher than the conventional C-TAILS. A systematic data analysis revealed that the optimized method is unbiased to amino acid composition, more reproducible and with higher MASCOT scores. Moreover, the introduction of SCII (Single-Charge Ion Inclusion) method to alleviate the charge deficiency of small peptides allowed an additional 26 % increase in identification number. With the optimized method, we identified 481 C-terminal peptides corresponding to 369 C-termini in E. Coli in a triplicate experiments using 80 μg each. Our optimized method would benefit the deep screening of C-terminomics and possibly help discover some novel C-terminal modifications.
HostingRepository	PRIDE
AnnounceDate	2015-09-15
AnnouncementXML	Submission_2015-09-15_02:37:00.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	yang zhang
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	monohydroxylated residue; acetylated residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2015-06-22 07:26:32	ID requested
⏵ 1	2015-09-15 02:37:02	announced

Zhang Y, He Q, Ye J, Li Y, Huang L, Li Q, Huang J, Lu J, Zhang X, Systematic Optimization of C-Terminal Amine-Based Isotope Labeling of Substrates Approach for Deep Screening of C-Terminome. Anal Chem, 87(20):10354-61(2015) [pubmed]

curator keyword: Technical

submitter keyword: Protein C-termini, acetylation, SCII

xumin zhang
contact affiliation	State Key Laboratory of Genetic Engineering, Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200438, China
contact email	xumin_zhang@fudan.edu.cn
lab head
yang zhang
contact affiliation	State Key Laboratory of Genetic Engineering, Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200438, China
contact email	wdyangzh@hotmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD002409
     1. Label: PRIDE project
     2. Name: Systematic optimization of C-TAILS approach for deep screening of C-terminomics