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

Johnson J, Harman VM, Franco C, Emmott E, Rockliffe N, Sun Y, Liu LN, Takemori A, Takemori N, Beynon RJ, la carte QconCAT protein standards for multiplexed quantification of user-specified target proteins. BMC Biol, 19(1):195(2021) [pubmed]

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