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Title: Evidence for two distinct intracellular pools of inorganic sulfate in Penicillium notatum

Journal Article · · J. Bacteriol.; (United States)
OSTI ID:5094022

A strain of Penicillium notatum unable to metabolize inorganic sulfate can accumulate sulfate internally to an apparent equilibrium concentration 10/sup 5/ times greater than that remaining in the medium. The apparent K/sub eq/ is near constant at all initial external sulfate concentrations below that which would eventually exceed the internal capacity of the cells. Under equilibrium conditions of zero net flux, external /sup 35/SO/sub 4//sup 2 -/ exchanges with internal, unlabeled SO/sub 4//sup 2 -/ at a rate consistent with the kinetic constants with the sulfate transport system. Efflux experiments demonstrated that sulfate occupies two distinct intracellular pools. Pool 1 is characterized by the rapid release of /sup 35/SO/sub 4//sup 2 -/ when the suspension of preloaded cells is adjusted to 10 mM azide at pH 8.4 (t/sub 1/2/, 0.38 min). /sup 35/SO/sub 4//sup 2 -/ in pool 1 also rapidly exchanges with unlabeled medium sulfate. Pool 2 is characterized by the slow release of /sup 35/SO/sub 4//sup 2 -/ induced by azide at pH 8.4 or unlabeled sulfate (t/sub 1/2/, 32 to 49 min). Early in the /sup 35/SO/sub 4//sup 2 -/ accumulation process, up to 78% of the total transported substrate is found in pool 1. At equilibrium, pool 1 accounts for only about 2% of the total accumulated /sup 35/SO/sub 4//sup 2 -/. Monensin (33 ..mu..m) accelerates the transfer of /sup 35/SO/sub 4//sup 2 -/ from pool 1 to pool 2. Valinomycin (0.2 ..mu..M) and tetraphynylboron/sup -/ (1 mM) retard the transfer of /sup 35/SO/sub 4//sup 2 -/ from pool 1 to pool 2. Pool 2 may reside in a vacuole or other intracellular organelle. A model for the transfer of sulfate from pool 1 to pool 2 is presented.

Research Organization:
Univ. of California, Davis
OSTI ID:
5094022
Journal Information:
J. Bacteriol.; (United States), Vol. 162:3
Country of Publication:
United States
Language:
English