Algal proteomics

Title: Changes in the proteome of green microalgae under abiotic stress conditions

Supervisor:  dr hab. Grzegorz Chrzanowski, prof. UR; e-mail: gchrzanowski@ur.edu.pl

Short description:

Environmental factors such as biotic and abiotic stresses can cause constraints growth and development of plants and green microalgae. These stresses also disturb cellular homeostasis, and consequently, the response is initiated to alleviate the impact of stress. The study of proteomes provides new insight into specific responses of photosynthetic organisms on proteins synthesis under stress conditions. On the other hand, increasing evidence from functional genomic studies suggests that information from transcriptomic data does not necessarily match the actual cellular protein complement, since different modifications may affect gene products, including post-transcriptional, co-translational, and degradative ones.

The aim of the present proposal is to give a critical account of the effects of stress conditions on protein composition and their role in response to abiotic stressors. Experiments will be conducted on the model green alga belonging to Chlorophyceae or Trebouxiophyceae class obtained from the algal bank.

We will also comparatively assess the influence of different solubilization buffer systems containing either a single-step process with a buffer containing one detergent (sodium dodecyl sulfate (SDS), Triton-X-100 (TRIT), no detergent, or a two-step solubilization process. Proteins will be analyzed either by PAGE electrophoresis after SPE fractionation with C18 columns or 2DE followed by LC–MS/MS. The LC–MS/MS analysis will be performed using the nanoACQUITY (Waters) system, coupled to an amazon ETD Ion Trap Mass Spectrometer (Bruker) after trypsin digestion.

Competences / skills and expectations:

The proposed experiments will aim to develop the ability to perform proteomic analysis using different electrophoretic methods combined with mass spectrometry, particularly two-dimensional electrophoresis.