Studies of protein-protein interaction using countercurrent distribution in aqueous two-phase systems. Partition behaviour of six Calvin-cycle enzymes from a crude spinach (Spinacia oleracea) chloroplast extract

Abstract

The partition behaviour of six enzymes of the Calvin cycle in extracts of chloroplasts from spinach (Spinacia oleracea) between two aqueous phases has been studied by countercurrent distribution. The enzymes showed distribution patterns which indicate heterogeneity and the presence of two or three fractions of most of the enzymes. When two of the enzymes, phosphoglycerate kinase and fructose-bisphosphate aldolase, were partitioned in both purified and partially purified form, they behaved like homogeneous substances. These results indicate that countercurrent distribution of crude extracts in aqueous two-phase systems is a useful method to study protein-protein interaction.

CP12 provides a new mode of light regulation of Calvin cycle activity in higher plants

“…GAPDH was also shown to be associated with PRK in a 600-kDa protein complex, made up by the A2B2 and the A4 forms of GAPDH, together with one PRK dimer (8). Further data suggest that GAPDH and PRK could be associated in a complex together with Rubisco (9). These three enzymes together were then proposed to be members of a 600-kDa multienzyme complex, in addition of ribose-5-P isomerase, and 3-phosphoglycerate kinase (PGK) (10,11).…”

Restoration of photosystem II photochemistry and carbon assimilation and related changes in chlorophyll and protein contents during the rehydration of desiccated Xerophyta scabrida leaves

“…On the other hand, in stressed Lemna minor fronds Rubisco was not degraded but gradually became polymerized to inactive aggregates, accompanied by a reduction in the number of sulphydryl groups ( Ferreira and Shaw, 1989 ). The Benson–Calvin cycle enzymes have a tendency to form soluble and membrane-bound multienzyme complexes ( Sainis and Harris, 1986 ; Gontero et al , 1988 , 1993 ; Sainis et al , 1989 ; Persson and Johansson, 1989 ; Hermoso et al , 1992 ; Anderson et al , 1995 ; Agarwal et al , 2009 ) with higher catalytic efficiency and less susceptibility to auto-oxidation and proteolysis than free enzymes ( Gontero et al , 1988 , 1993 ).…”

Enzyme-enzyme interaction in the chloroplast: Glyceraldehyde-3-phosphate dehydrogenase, triose phosphate isomerase and aldolase

“…Persson and Johansson (1989) found evidence of interaction between each of these enzymes and other stromal components, but they did not identify the interacting species. The evidence for interaction between mammalian glyceraldehyde-3-P dehydrogenase and aldolase includes effects on fluorescence intensity when the enzymes are mixed (Neuzil et al 1990), kinetic experiments (Ov~idi and Keleti 1978;Kvassman et al 1988) and isotope-dilution experiments (Orosz and Ovddi 1987).…”

“…The experiments of Mtiller (1972), Gontero et al (1988), Persson and Johansson (1989), Macioszek et al (1990), Skrukrud et al (1991), Stiss et al (1993), Hosur et al (1993) and Anderson et al (1993) suggest that there is interaction between the five Calvin-cycle enzymes which convert ribose-5-phosphate to glyceraldehyde-3-E The experiments described here indicate that L.E. Anderson et al: Enzyme-enzyme interaction in the chloroplast glyceraldehyde-3-P dehydrogenase, aldolase and triose-P isomerase interact within the chloroplast.…”