PXD039306

PXD039306 is an original dataset announced via ProteomeXchange.

Title	Evolutionary and cellular analysis of the'dark'pseudokinase PSKH2
Description	Pseudokinases, so named because they lack one or more conserved canonical amino acids that define their catalytically-active relatives, have evolved a variety of biological functions in both prokaryotic and eukaryotic organisms. Human PSKH2 is closely related to the canonical kinase PSKH1, which maps to the CAMK family of protein kinases. Primates encode PSKH2 in the form of a pseudokinase, which is predicted to be catalytically inactive due to loss of the invariant catalytic Asp residue. Although the biological role(s) of vertebrate PSKH2 proteins remains unclear, we previously identified species-level adaptions in PSKH2 that have led to the appearance of kinase or pseudokinase variants in vertebrate genomes alongside a canonical PSKH1 paralog. In this paper we confirm that, as predicted, PSKH2 lacks detectable protein phosphotransferase activity, and exploit structural informatics, biochemistry and cellular proteomics to begin to characterise vertebrate PSKH2 orthologues. AlphaFold 2-based structural analysis predicts functional roles for both the PSKH2 N- and Cregions that flank the pseudokinase domain core, and cellular truncation analysis confirms that the N-terminal domain, which contains a conserved myristoylation site, is required for both stable human PSKH2 expression and localisation to a membrane-rich subcellular fraction containing mitochondrial proteins. Using mass spectrometry-based proteomics, we confirm that human PSKH2 is part of a cellular mitochondrial protein network, and that its expression is regulated through client-status within the HSP90/Cdc37 molecular chaperone system. HSP90 interactions are mediated through binding to the PSKH2 C-terminal tail, leading us to predict that this region might act as both a cis and trans regulatory element, driving outputs linked to the PSKH2 pseudokinase domain that are important for functional signalling.
HostingRepository	PRIDE
AnnounceDate	2023-03-11
AnnouncementXML	Submission_2023-03-11_11:23:38.378.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD039306
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	leonarddaly
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-01-06 06:57:29	ID requested
⏵ 1	2023-03-11 11:23:39	announced

10.6019/PXD039306;

Byrne DP, Shrestha S, Daly LA, Marensi V, Ramakrishnan K, Eyers CE, Kannan N, Eyers PA, Evolutionary and cellular analysis of the 'dark' pseudokinase PSKH2. Biochem J, 480(2):141-160(2023) [pubmed]

submitter keyword: Pseudokinase, PSKH2

Claire EEyers
contact affiliation	Centre for Proteome Research, Biochemistry and Systems Biology Department, University of Liverpool
contact email	ceyers@liverpool.ac.uk
lab head
leonarddaly
contact affiliation	University of Liverpool
contact email	leonard.daly@liverpool.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD039306
     1. Label: PRIDE project
     2. Name: Evolutionary and cellular analysis of the'dark'pseudokinase PSKH2