Rubisco (Ribulose 1, 5-bisphosphate carboxylase/oxygenase, EC 4.1.1.39) is crucial in biological circumstance fluctuation. Although disassembly of Rubisco after chill treatment has been reported in previous studies, there is only little known data on Rubisco interactive proteins involved in the disassembly process of Rubisco. Both repression of net photosynthesis rate and disassembly of Rubisco large subunits (Ru-L) have been investigated in the wild type, Arabidopsis thaliana (Col-0), treated at 4℃ for 4 h and 24 h together with their 24 h recoveries at 20℃. Co-immunoprecipitation coupled with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and MALDI-TOF MS identification was used to explore Rubisco-interacting proteins. Five protein candidates were profiled. The identified AAA-type ATPase family and glycosyltransferase were determined crucial for Rubisco activity. It is also strongly correlated to cold acclimation. Results suggest that the disassembly of Rubisco might have been the main cause of photosynthesis rate reduction under chill conditions, rather than photosystem or biogenesis involvement.