The current model of the Ca²⁺ homeostasis in yeast considers three transporters which mediate calcium efflux from cytoplasm, namely Golgi (Pmr1p) and vacuolar (Pmc1p) Ca²⁺-ATPases and vacuolar Ca²⁺/H⁺ exchanger (Vcx1p). A role of the ER, plasma membrane (PM) and their Ca²⁺ transporters is still controversial or even ignored. The published data on the effective maintenance of the low Ca²⁺ in the yeast cytosol in vivo by Vcx1p are in contradiction with widely accepted opinion on its low affinity for Ca²⁺. We show here that extra-cellular glucose stimulated the Ca²⁺/H⁺ exchangers of total membranes (TM) more than 7 times and Ca²⁺-ATPases 2 times leading to 80% contribution of the exchangers in the Ca²⁺ efflux from cytosol. The Ca²⁺/H⁺ exchangers of TM exhibited the same order of affinity for Ca²⁺ and higher velocity of the transport in comparison with Ca²⁺-ATPases (K_M=100 nM; V_max=4.3 nmol/min x mg after the stimulation with glucose and K_M=60 nM and V_max=0.52 nmol/min x mg before activation). Ca²⁺-ATPases of TM have K_M= 50 nM and 46 nM with V_max=1.25 and 0.28 nmol/min x mg, after and before the activation with glucose, respectively. The fractionation of membranes in sucrose density gradient has revealed that different organelles of the secretory pathway possess both type of Ca²⁺ transporters showing a tendency of their selective regulation by extra-cellular glucose and higher activity of the exchanger. All compartments of secretory pathway participate in Ca²⁺ homeostasis with a remarkable contribution of the ER/PM in Ca²⁺ efflux from cytosol (26%). These data together with the finding of the high affinity and high activity of the Ca²⁺/H⁺ exchangers in the organelles of the secretory pathway ensure and explain their important role in the Ca²⁺ homeostasis and Ca²⁺ signaling.