PXD020104-1

PXD020104 is an original dataset announced via ProteomeXchange.

Title	Generation and validation of recombinant antibodies to study tRNA synthetases in health and disease
Description	Aminoacyl tRNA synthetases (aaRSs) have long been viewed as mere housekeeping proteins and have therefore often been overlooked in drug discovery. However, recent findings have revealed that many aaRSs have non-canonical functions and several of them have been linked to autoimmune diseases, cancer and neurological disorders. In order to study these proteins further, recombinant high-affinity antibodies have been generated to a selection of thirteen cytoplasmic and one mitochondrial aaRSs. Selected domains of these proteins were produced recombinantly in Escherichia coli and used as antigens in phage display selections using a synthetic human single-chain fragment variable (scFv) library. All targets yielded large sets of antibody candidates that were validated through a panel of binding assays against the purified antigen. Furthermore, the top performing binders were tested in immunoprecipitation followed by mass spectrometry (IP-MS) for their ability to capture the endogenous protein from mammalian cell lysates. Interestingly, for antibodies targeting individual members of the multi-tRNA synthetase complex (MSC), we were able to detect all members of the complex, co-immunoprecipitating with the target, in several cell types. The functionality of a sub-set of binders for each target was also confirmed in immunofluorescence. To conclude, we have created an open source resource, in the form of high quality recombinant proteins and antibodies to accelerate and empower future research of the role of aaRSs in health and disease.
HostingRepository	PRIDE
AnnounceDate	2020-08-13
AnnouncementXML	Submission_2020-08-12_22:14:38.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Susanne Gräslund
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2020-06-29 07:32:25	ID requested
⏵ 1	2020-08-12 22:14:38	announced
2	2024-10-22 05:10:39	announced	2024-10-22: Updated project metadata.

Dataset with its publication pending

submitter keyword: aminoacyl tRNA synthetase, phage display, recombinant antibodies, antibody validation

Susanne Gräslund
contact affiliation	Structural Genomics Consortium, Dept. of Medicine, Division of Rheumatology, CMM, L8:02, Karolinska Institutet, 171 76 Solna, Sweden
contact email	susanne.graslund@ki.se
lab head
Susanne Gräslund
contact affiliation	Structural Genomics Consortium, Department of Medicine Solna, Karolinska Institutet, CMM L8:02, 171 76 Solna, Sweden
contact email	susanne.graslund@ki.se
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD020104
     1. Label: PRIDE project
     2. Name: Generation and validation of recombinant antibodies to study tRNA synthetases in health and disease