Low-affinity potassium uptake system in the archaeon Methanobacterium thermoautotrophicum: Overproduction of a 31-kilodalton membrane protein during growth on low-potassium medium

Autor(en): Glasemacher, J
Siebers, A
Altendorf, K 
Schonheit, P
Stichwörter: BACILLUS-ACIDOCALDARIUS; CATIONS; ESCHERICHIA-COLI; K+; METHANOGENIC BACTERIA; METHANOSPIRILLUM-HUNGATEI; Microbiology; PURIFICATION; TRANSLOCATING KDP-ATPASE; TRANSPORT
Erscheinungsdatum: 1996
Herausgeber: AMER SOC MICROBIOLOGY
Journal: JOURNAL OF BACTERIOLOGY
Volumen: 178
Ausgabe: 3
Startseite: 728
Seitenende: 734
Zusammenfassung: 
During growth on low-K+ medium (1 mM K+), Methanobacterium thermoautotrophicum accumulated K+ up to concentration gradients ([K+](intracellular)/[K+](extracellular)) of 25,000- to 50,000-fold. At these gradients ([K+](extracellular) Of < 20 mu M), growth ceased but could be reinitiated by the addition of K+ or Rb+. During K+ starvation, the levels of a protein with an apparent molecular weight of 31,000 increased about sixfold. The protein was associated with the membrane and could be extracted by detergents. Cell suspensions of M. thermoautotrophicum obtained after K+-limited growth catalyzed the transport of both K+ and Rb+ with apparent K-m and V-max values of 0.13 mM and 140 nmol/min/mg, respectively, for K+ and 3.4 mM and 140 nmol/min/mg, respectively, for Rb+. Rb+ competitively inhibited K+ uptake with an inhibitor constant of about 10 mM. Membranes of K+-starved cells did not exhibit Kt-stimulated ATPase activity. Immunoblotting with antisera against Escherichia coli Kdp-ATPase did not reveal any specific cross-reactivity against membrane proteins of K+-starved cells. Cells of M. thermoautotrophicum grown at a high potassium concentration (50 mM) catalyzed K+ and Rb+ transport at similar apparent K-m values (0.13 mM for K+ and 3.3 mM for Rb+) but at significantly lower apparent V-max values (about 60 nmol/min/mg for both K+ and Rb+) compared with K+-starved cells. From these data, it is concluded that the archaeon M. thermoautotrophicum contains a low-affinity K+ uptake system which is overproduced during growth on low-K+ medium.
ISSN: 00219193
DOI: 10.1128/jb.178.3.728-734.1996

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