PXD003324

PXD003324 is an original dataset announced via ProteomeXchange.

Title	Multiplex selected reaction monitoring assay for quantification of 15 human tissue kallikreins in biological samples
Description	Human tissue kallikreins (KLKs) are a group of 15 secreted serine proteases encoded by the largest contiguous cluster of protease genes in the human genome. KLKs are involved in coordination of numerous physiological functions including regulation of blood pressure, neuronal plasticity, skin desquamation and semen liquefaction, and thus represent promising diagnostic and therapeutic targets. Until now, quantification of KLKs in biological and clinical samples was accomplished only by enzyme-linked immunosorbent assays (ELISA). In this work, we developed a multiplex selected reaction monitoring (SRM) assay for the simultaneous quantification of all 15 KLKs. Proteotypic peptides for each KLK were carefully selected based on experimental data and then multiplex in a single assay. Assay performance was evaluated using three different mass spectrometry platforms including triple quadrupole, quadrupole-ion trap and quadrupole-orbitrap instruments. Heavy isotope-labeled synthetic peptides with a quantifying tag were used for absolute quantification of KLKs in seminal plasma, sweat and cervico-vaginal fluid, with limits of detection ranging from 5 to 500 ng/mL. Analytical performance of the SRM assay was evaluated by measuring endogenous KLKs in relevant clinical samples and results were compared to selected ELISAs. The multiplex SRM assay was proven to be an accurate, reproducible, sensitive and highly specific alternative to the existing antibody-based assays. Finally, we used immunoenrichment-SRM and ELISA to unambiguously detect and quantify seminal plasma levels of kallikrein-4, a highly prostate-specific protein and a potential biomarker of prostate-related diseases. The presented multiplex SRM assay is an alternative analytical tool to study the biological and pathological roles of human KLKs.
HostingRepository	PRIDE
AnnounceDate	2016-07-08
AnnouncementXML	Submission_2016-07-08_04:52:57.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Theano Karakosta
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	deamidated residue; monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2015-12-11 01:59:50	ID requested
⏵ 1	2016-07-08 04:52:58	announced

Karakosta TD, Soosaipillai A, Diamandis EP, Batruch I, Drabovich AP, Quantification of Human Kallikrein-Related Peptidases in Biological Fluids by Multiplatform Targeted Mass Spectrometry Assays. Mol Cell Proteomics, 15(9):2863-76(2016) [pubmed]

curator keyword: Biomedical

submitter keyword: Kallikreins, mass spectrometry, selected reaction monitoring, seminal plasma, cervicovaginal fluid, sweat, KLK4 secretion

Andrei P. Drabovich
contact affiliation	Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada
contact email	andrei.drabovich@utoronto.ca
lab head
Theano Karakosta
contact affiliation	University of Toronto
contact email	theano.karakosta@utoronto.ca
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD003324
     1. Label: PRIDE project
     2. Name: Multiplex selected reaction monitoring assay for quantification of 15 human tissue kallikreins in biological samples