PXD023073-1

PXD023073 is an original dataset announced via ProteomeXchange.

Title	NeissLock: an inducible protein anhydride for covalent targeting of endogenous proteins
Description	Effective and convenient covalent protein conjugation could benefit diverse biological systems, stabilizing key protein protein interactions which are otherwise reversible. Neisseria meningitidis contains a protein which undergoes autoproteolysis via an anhydride. Here, we harness this spontaneously generated electrophile for covalent targeting of unmodified endogenous proteins. In ‘NeissLock’, a binding protein genetically fused to the self-processing module (SPM) docks to its target protein. Upon triggering with calcium, the aspartic anhydride generated at the C-terminus of the binding protein allows nucleophilic attack by nearby residues on the target protein, so ligating the proteins. We established a computational tool to search the Protein Data Bank, assessing proximity of amines to C-termini. We validated and optimized the NeissLock concept using the Ornithine Decarboxylase/Antizyme complex. A range of nucleophiles on the target (α-amine or ε-amines) could rapidly react with the anhydride, but reaction was blocked if the partner protein did not dock. Surprisingly, the optimal pH for covalent ligation was 7.0. We then armed Transforming Growth Factor-α with SPM and established covalent targeting to Epidermal Growth Factor Receptor at the surface of living cells. NeissLock harnesses exceptional protein chemistry to allow covalent targeting of endogenous proteins under mild conditions with up to 80% yield, allowing new possibilities for molecular engineering.
HostingRepository	PRIDE
AnnounceDate	2021-02-01
AnnouncementXML	Submission_2021-01-31_22:25:35.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Shabaz Mohammed
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2020-12-11 07:03:53	ID requested
⏵ 1	2021-01-31 22:25:36	announced

Scheu AHA, Lim SYT, Metzner FJ, Mohammed S, Howarth M, NeissLock provides an inducible protein anhydride for covalent targeting of endogenous proteins. Nat Commun, 12(1):717(2021) [pubmed]

submitter keyword: Protein engineering, Protein conjugation, Proximity labeling, Synthetic biology, Chemical biology, Nanoassembly

Mark Howarth
contact affiliation	Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, 7 UK.
contact email	mark.howarth@bioch.ox.ac.uk
lab head
Shabaz Mohammed
contact affiliation	University of Oxford
contact email	shabaz.mohammed@chem.ox.ac.uk
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/02/PXD023073

PRIDE project URI

• PRIDE
  1. PXD023073
     1. Label: PRIDE project
     2. Name: NeissLock: an inducible protein anhydride for covalent targeting of endogenous proteins