Phosphatidylglycerol (PG) is the only anionic phospholipid in photosynthetic membrane and the important component of photosystem I (PSI). In this study, the interaction of PG with PSI particle from spinach was investigated by using reconstitution method. The results from the properties of electron transport, fluorescence emission, turbidity, and protein secondary structures in PSI complex incorporated into PG liposomes revealed the existence of PSI-PG interactions. A stimulation and an inhibition of oxygen uptake in PSI particle at a low and higher PG/chlorophyll mass ratio, respectively, were observed. Moreover, an additional enhancement and depression of electron flow in the PSI-PG complexes were occurred in the reaction medium containing CaCl2 at concentrations below and above 5 mM, the aggregation threshold of the reconstituted membranes, respectively. The results demonstrated that the maintenance of the structural optimization was needed for a stimulation of electron transport at a low PG/PSI mass ratio, while a decay of this PSI activity at high PG/PSI ratio was the result of inhibition of the energy transfer from LHCI to PSI reaction center induced by the dissociation of LHCI-680.