Anti-CURT1A | Curvature thylakoid 1A

Hordeum vulgare, Phaseolus coccineus

Plant species: Arabidopsis LD – Arabidopsis thaliana grown under long day conditions Arabidopsis SD – Arabidopsis thaliana grown under short day conditions Runner bean – Phaseolus coccineus L. Pea – Pisum sativum L. Experimental procedure: Samples of isolated thylakoids containing 50 µg of chlorophyll were denatured with 150 mL double diluted Roti®-Load 1 (ROTH, Art.-Nr. K929.1) at 95 °C for 2 min. Denatured samples containing 0.5 and 1 µg of chlorophyll were loaded in the gel wells and separated on 14% SDS-PAGE gels and blotted for 45 min at 105 V to PVDF membrane using wet transfer. Blot was blocked with 5% Amersham™ ECL Prime Blocking Agent (cat no: RPN418) in TBS-T for 30 min at room temperature (RT) with agitation. The blot was incubated with the primary antibody at a dilution of 1:1000 overnight at 4˚C with agitation. The antibody solution was decanted and the blot was washed 2 times for 5 min in TBS-T at RT with agitation. The blot was incubated using a secondary antibody (goat anti-rabbit IgG HRP conjugated, from Agrisera, AS09 602) diluted to 1: 25000 in 1% Blocking Agent in TBS-T for 1h at RT with agitation. The blot was washed 2 times for 5 min in TBS-T and developed for 5 min with chemiluminescent reagent, according to instructions. Exposure time was 12 minutes in C-Digit chemiluminescence scanner (Li-COR).

Dr Radosław Mazur, Faculty of Biology, University of Warsaw, Poland

Curvature thylakoid 1A (CURT1A) belongs to a protein family, conserved in plants and cyanobaceria. There are four Arabidopsis thaliana CURT1 proteins: CURT1A,B,C and D.  It is proposed that CURT1 proteins modify thylakoid architecture by inducing membrane curvature at grana margins.