PXD027047

PXD027047 is an original dataset announced via ProteomeXchange.

Title	Synthetic biology meets proteomics: Construction of à la carte QconCATs for absolute protein quantification
Description	We report a new approach to the assembly and construction of QconCATs, quantitative concatamers for proteomic applications that yield stoichiometric quantities of sets of stable isotope-labelled internal standards. The new approach is based on synthetic biology precepts of biobricks, making use of loop assembly to construct larger entities from individual biobricks. It offers a major gain in flexibility of QconCAT implementation and enables rapid and efficient editability that permits, for example, substitution of one peptide for another. The basic building block (a Qbrick) is a segment of DNA that encodes two or more quantification peptides for a single protein, readily held in a repository as a library resource. These Qbricks are then assembled in a one tube ligation reaction that enforces the order of assembly, to yield short QconCATs that are useable for small quantification products. However, the DNA context of the short also allows a second cycle of assembly such that five different short QconCATs can be assembled into a longer QconCAT in a second, single tube ligation. From a library of Qbricks, a bespoke QconCAT can be assembled quickly and efficiently in a form suitable for expression and labelling in vivo or in vitro. We refer to this approach as the ALACAT strategy as it permits à la carte design of quantification standards.
HostingRepository	PanoramaPublic
AnnounceDate	2022-07-18
AnnouncementXML	Submission_2022-07-18_22:24:52.301.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	supported by repository but incomplete data and/or metadata
PrimarySubmitter	Nobuaki Takemori
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	Label:13C(6)15N(2); Label:13C(6)15N(4)
Instrument	QTRAP 5500

Revision	Datetime	Status	ChangeLog Entry
0	2021-07-01 16:03:24	ID requested
⏵ 1	2022-07-18 22:24:53	announced

submitter keyword: ALACAT, QconCAT, Cell-free protein synthesis

Nobuaki Takemori
contact affiliation	Ehime University
contact email	takemori@m.ehime-u.ac.jp
lab head
Nobuaki Takemori
contact affiliation	Ehime University
contact email	takemori@m.ehime-u.ac.jp
dataset submitter

Panorama Public dataset URI