Proteomic analysis of small heat shock protein isoforms in barley shoots.
The analysis of stress-responsiveness in plants is an important route to the discovery of genes conferring stress tolerance and their use in breeding programs. High temperature is one of the environmental stress factors that can affect the growth and quality characteristics of barley (Hordeum vulgare). In this study a proteomic analysis (2D-PAGE, MS) was used to detect the effects of heat shock on the protein pattern of an abiotic stress-tolerant (Mandolina) and an abiotic stress-susceptible (Jubilant) barley cultivar. Evaluation of two-dimensional gels revealed several proteins to be differentially expressed as a result of heat stress in both cultivars. The protein spots of interest were, after an in-gel tryptic digestion, further investigated by mass spectrometry. For the analysis of the peptide mixture, we both used a matrix-assisted laser desorption/ionization (MALDI) tandem time of flight mass spectrometer (TOF/TOF) and an automated nano-HPLC system coupled to an electrospray ionization-quadrupole linear ion trap (Q-TRAP) instrument. The hyphenation of the latter techniques proved to be a powerful technique as shown by the identification of six isoforms of a 16.9 kDa sHSP in one single spot. We observed that S-adenosylmethionine synthetase (SAM-S) was differentially expressed between the two cultivars. Recent results refer to the role of SAM-S as being involved in abiotic stress tolerance. Furthermore, comparison of the heat shock treated samples also revealed several small heat shock proteins (sHSP), of which distinct isoforms could be characterised.
Faculty of Food Science, Budapest University of Economic Sciences and Public Administration, Villányi 29-43, H-1118 Budapest, Hungary.
A
A
Süle
F
F
Vanrobaeys
Gy
G
Hajós
J
J
Van Beeumen
B
B
Devreese
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D015870:Gene Expression; D006360:Heat-Shock Proteins / Q000032:analysis*; D001467:Hordeum / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D008970:Molecular Weight; D018520:Plant Shoots / Q000378:metabolism*; D020033:Protein Isoforms; D020543:Proteome / Q000032:analysis; D012436:S-Adenosylmethionine / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D006360:Heat-Shock Proteins; D020033:Protein Isoforms; D020543:Proteome; D012436:S-Adenosylmethionine