Comparison of lactose uptake in resting and energized Escherichia coli cells: high rates of respiration inactivate the lac carrier.
نویسندگان
چکیده
The transport of lactose by Escherichia coli cells was radically different in the absence and in the presence of an exogenous energy source: in the former case, the time course of lactose accumulation was monotonous; in the latter case, lactose accumulation reached a maximum and then decreased to a final steady-state level lower than that observed in the absence of an energy source. We show that this "overshoot" is the result of a decrease in the influx rate and of an increase in the rate constant of efflux as lactose accumulates. These phenomena were irreversible. The extent of the overshoot was dependent upon the experimental conditions: it was maximal at alkaline pH, for low external potassium concentrations, and for relatively high external lactose concentrations (around or above the KT of uptake). The addition of an energy source to resting E. coli cells resulted in an increase in both the electrochemical gradient of protons and in the rate of respiration. We demonstrate that the overshoot is the result of the latter and unrelated to the former. We observed an irreversible decrease in the membrane potential as lactose accumulated in the presence of an exogenous energy source. We discuss the whole of our data in terms of an irreversible inactivation of the lactose carrier as a result of a possible interaction with the respiratory chain.
منابع مشابه
CONSTRUCTION OF RECOMBINANT PLASMIDS FOR PERIPLASMIC EXPRESSION OF HUMAN GROWTH HORMONE IN ESCHERICHIA COLI UNDER T7 AND LAC PROMOTERS
In order to study the periplasmic expression of human growth hormone (hGH) in Escherichia coli, the related cDNA was inserted in two expression plasmids carrying pelB signal peptide, one with lac bacterial promoter and the other with a bacteriophage T7-based promoter. The recombinant plasmids were moved to TG1 and BL21 strains of E. coli, respectively. To induce the expression systems, IPTG and...
متن کاملComparison of T7- and Lac-Based Systems for the Periplasmic Expression of Human Granulocyte Macrophage Colony Stimulating Factor in Escherichia coli
متن کامل
Preparation, characterization, and properties of monoclonal antibodies against the lac carrier protein from Escherichia coli.
Monoclonal antibodies directed against the lac carrier protein purified from the membrane of Escherichia coli were prepared by somatic cell fusion of mouse myeloma cells with splenocytes from an immunized mouse. Several clones produce antibodies that react with the purified protein as demonstrated by solid-phase radioimmunoassay and by immunoblotting experiments; culture supernatants from the c...
متن کاملNonadaptive mutations occur on the F' episome during adaptive mutation conditions in Escherichia coli.
One of the most studied examples of adaptive mutation is a strain of Escherichia coli, FC40, that cannot utilize lactose (Lac-) but that readily reverts to lactose utilization (Lac+) when lactose is its sole carbon source. Adaptive reversion to Lac+ occurs at a high rate when the Lac- allele is on an F' episome and conjugal functions are expressed. It was previously shown that nonselected mutat...
متن کاملPopulation dynamics of a Lac- strain of Escherichia coli during selection for lactose utilization.
During selection for lactose utilization, Lac+ revertants of FC40, a Lac- strain of Escherichia coli, appear at a high rate. Yet, no Lac+ revertants appear in the absence of lactose, or in its presence if the cells have another, unfulfilled requirement for growth. This study investigates more fully the population dynamics of FC40 when incubated in the absence of a carbon source or when undergoi...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of bacteriology
دوره 154 1 شماره
صفحات -
تاریخ انتشار 1983