Identification of Reference and Biomarker Proteins in Chlamydomonas reinhardtii Cultured under Different Stress Conditions.

Jianan Shi,Yalu Guo,Teng Huang,Liyun Li,Guozhen Liu,Jian Wei,Shuaijie Chai,Shijuan Dou

doi:10.3390/ijms18081822

Abstract

Reference proteins and biomarkers are important for the quantitative evaluation of protein abundance. Chlamydomonas reinhardtii was grown under five stress conditions (dark, cold, heat, salt, and glucose supplementation), and the OD750 and total protein contents were evaluated on days 0, 1, 2, 4, and 6 of culture. Antibodies for 20 candidate proteins were generated, and the protein expression patterns were examined by western blotting. Reference protein(s) for each treatment were identified by calculating the Pearson’s correlation coefficient (PCC) between target protein abundance and total protein content. Histone H3, beta tubulin 1 (TUB-1), ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (RBCL), and mitochondrial F1F0 ATP synthase subunit 6 (ATPs-6) were the top reference proteins, because they were expressed stably under multiple stress conditions. The average relative-fold change (ARF) value of each protein was calculated to identify biomarkers. Heat shock protein 90B (HSP90B), flagellar associated protein (FAP127) and ATP synthase CF0 A subunit (ATPs-A) were suitable biomarkers for multiple treatments, while receptor of activated protein kinase C1 (RCK1), biotin carboxylase (BCR1), mitochondrial phosphate carrier protein (MPC1), and rubisco large subunit N-methyltransferase (RMT1) were suitable biomarkers for the dark, cold, heat, and glucose treatments, respectively.

Highlights

Microalgae are photosynthetic microorganisms that are used in a wide variety of applications
GTrohwetmh oofrpCh. roelionghyaradptipi eCaerllasnucnedoefr CSt.rreesisnhardtii cells cultured under different stress conditions is showTnheinmFoirgpuhroelo1gAy, aapnpdeathraencceellofnCu.mrebinerh,arOdDtii7c50e,llasncdulOtuDre6d80 ugnrodwerthdicffuerevnetssatresshcowndnitinonFsiigsusrheo1wBn– iDn,Frigesuprec1tAiv,ealnyd. the cell number, optical density at 750 nm (OD750), and OD680 growth curves are shown in Figure 1B–D, respectively
C. reinhardtii was cultivated under five stress conditions: dark, cold, heat, salt, and glucose supplementation

Summary

Introduction

Microalgae are photosynthetic microorganisms that are used in a wide variety of applications Due to their relatively high lipid/biogas productivity, microalgae are promising for biofuel production [1,2]. Microalgae can biosynthesize various pigments, antioxidants, β-carotenes, proteins, and vitamins, which can be used to produce high-value products. They can be grown on non-arable land and require less land than terrestrial crops; they neither compromise food production nor compete with agriculture [4]. These microorganisms can grow in a wide variety of environmental conditions, microalgae production still faces significant problems related to stress during the culture process

Methods

Results

Conclusion