A1 Refereed original research article in a scientific journal

Development of a Quantitative SRM-Based Proteomics Method to Study Iron Metabolism of Synechocystis sp. PCC 6803




AuthorsLinda Vuorijoki, Janne Isojärvi, Pauli Kallio, Petri Kouvonen, Eva-Mari Aro, Garry L. Corthals, Patrik R. Jones, Dorota Muth-Pawlak

PublisherACS Publications

Publication year2016

JournalJournal of Proteome Research

Journal acronymJPR

Volume15

Issue1

First page 266

Last page279

Number of pages14

ISSN1535-3893

DOIhttps://doi.org/10.1021/acs.jproteome.5b00800(external)


Abstract

The cyanobacterium Synechocystis sp. PCC 6803 (S. 6803) is a well-established model species in oxygenic photosynthesis research and a potential host for biotechnological applications. Despite recent advances in genome sequencing and microarray techniques applied in systems biology, quantitative proteomics approaches with corresponding accuracy and depth are scarce for S. 6803. In this study, we developed a protocol to screen changes in the expression of 106 proteins representing central metabolic pathways in S. 6803 with a targeted mass spectrometry method, selected reaction monitoring (SRM). We evaluated the response to the exposure of both short- and long-term iron deprivation. The experimental setup enabled the relative quantification of 96 proteins, with 87 and 92 proteins showing adjusted p-values <0.01 under short- and long-term iron deficiency, respectively. The high sensitivity of the SRM method for S. 6803 was demonstrated by providing quantitative data for altogether 64 proteins that previously could not be detected with the classical data-dependent MS approach under similar conditions. This highlights the effectiveness of SRM for quantification and extends the analytical capability to low-abundance proteins in unfractionated samples of S. 6803. The SRM assays and other generated information are now publicly available via PASSEL and Panorama.




Last updated on 2024-26-11 at 22:45