How Hemorrhagic E. coli Resists the Acid Environment pH Sensitivity of the Glu-GABA Antiporter 300 GadC of the Stomach Adic 250- Recent years have been marked by a series of food poisoning outbreaks involving hemorrhagic (producing internal bleeding) strains of the bacterium Escherichia coli (E. coli). Bacteria are often a source of food poisoning, typically milder infections caused by food-borne strepto- coccal bacteria. Less able to bear the extremely acidic conditions encountered by food in the human stomach (pH 2), E. coli has not been as common a problem. The hemorrhagic strains of E. coli responsible for recent out- breaks seem to have evolved more elaborate acid-resistance 200 150 100 50 5 6 7 systems. How do hemorrhagic E. coli bacteria survive in the acid environment of the stomach? The problem they face, pH in essence, is that they are ions, many of which diffuse into their cells. To rid them- selves of these excess hydrogen ions, the E. coli cells use a clever system to pump hydrogen ions back out of their cells First, the hemorrhagic E. coli cells take up cellular hydrogen ions by using the enzyme glutamic acid decarbox- ylase (GAD) to convert the amino acid glutamate to Y-aminobutyric acid (GABA), a decarboxylation reaction that consumes a hydrogen ion. Second, the hemorrhagic E. coli export this GABA from their cell cytoplasm using called GadC (this transmembrane protein channel is called an antiporter because it transports two molecules across the membrane in opposite directions). However, to survive elsewhere in the human body, it is important that the Glu-GABA antiporter of hemorrhagic E. coli not function, lest it short-circuit metabolism. To see if submerged in a sea of hydrogen Analysis 1. Applying Concepts a. Variable. In the graph, what is the dependent variable? b. Substrate. What is a substrate? In this investigation, what are the substrates that are accumulating? c. pH. What is the difference in hydrogen ion concentration between pH 5 and pH 7? How many times more (or less) is that? Explain. 2. Interpreting Data a. Does the amount of amino acid transported in the 10-minute experimental interval (expressed as substrate accumulation) vary with pH for the arginine-transporting AdiC antiporter? For the glutamate-transporting GadC antiporter? b. Compare the amount of substrate accumulated by AdiC in 10 minutes at pH 9.0 with that accumulated at pH 5.0. What fraction of the low pH activity is observed at the higher pH? c. In a similar fashion, compare the amount of substrate accumulated by GadC at pH 9.0 with that accumulated at pH 5.0. What fraction of the low pH activity is observed at the higher pH? a Glu-GABA antiporter the GadC antiporter indeed functions only in acid environ- ments, investigators compared its activity at a variety of pHs with that of a different amino acid antiporter called AdiC, which transports arginine out of cells under a broad range of conditions. The results of monitoring transport for 10 minutes are presented in the graph. Outside Inside cell cell 3. Making Inferences Would you say that the GadC antiporter exhibits the same pH dependence as the AdiC antiporter? If not, which antiporter is less active at nonacid pHs? GABA 4. Drawing Conclusions Is the glutamate-GABA antiporter GadC active at nonacid pHs? 5. Further Analysis The GadC antiporter also Glutamate transports the amino acid glutamine (Gln). Do you think this activity has any role to play in combating low pH environments? How would you test this hypothesis? Chapter 5 Membranes 111 Substrate accumulation (nmol per mg protein) Inquiry & Analysis

Curren'S Math For Meds: Dosages & Sol
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How Hemorrhagic E. coli
Resists the Acid Environment
pH Sensitivity of the Glu-GABA Antiporter
300
GadC
of the Stomach
Adic
250-
Recent years have been marked by a series of food
poisoning outbreaks involving hemorrhagic (producing
internal bleeding) strains of the bacterium Escherichia coli
(E. coli). Bacteria are often a source of food poisoning,
typically milder infections caused by food-borne strepto-
coccal bacteria. Less able to bear the extremely acidic
conditions encountered by food in the human stomach
(pH 2), E. coli has not been as common a problem. The
hemorrhagic strains of E. coli responsible for recent out-
breaks seem to have evolved more elaborate acid-resistance
200
150
100
50
5
6
7
systems.
How do hemorrhagic E. coli bacteria survive in the
acid environment of the stomach? The problem they face,
pH
in essence, is that they are
ions, many of which diffuse into their cells. To rid them-
selves of these excess hydrogen ions, the E. coli cells use a
clever system to pump hydrogen ions back out of their
cells
First, the hemorrhagic E. coli cells take up cellular
hydrogen ions by using the enzyme glutamic acid decarbox-
ylase (GAD) to convert the amino acid glutamate to
Y-aminobutyric acid (GABA), a decarboxylation reaction that
consumes a hydrogen ion.
Second, the hemorrhagic E. coli export this GABA
from their cell cytoplasm using
called GadC (this transmembrane protein channel is called
an antiporter because it transports two molecules across the
membrane in opposite directions).
However, to survive elsewhere in the human body, it
is important that the Glu-GABA antiporter of hemorrhagic
E. coli not function, lest it short-circuit metabolism. To see if
submerged in a sea of hydrogen
Analysis
1. Applying Concepts
a. Variable. In the graph, what is the dependent
variable?
b. Substrate. What is a substrate? In this
investigation, what are the substrates that are
accumulating?
c. pH. What is the difference in hydrogen
ion concentration between pH 5 and pH 7?
How many times more (or less) is that? Explain.
2. Interpreting Data
a. Does the amount of amino acid transported
in the 10-minute experimental interval
(expressed as substrate accumulation) vary
with pH for the arginine-transporting AdiC
antiporter? For the glutamate-transporting
GadC antiporter?
b. Compare the amount of substrate accumulated
by AdiC in 10 minutes at pH 9.0 with that
accumulated at pH 5.0. What fraction of the
low pH activity is observed at the higher pH?
c. In a similar fashion, compare the amount of
substrate accumulated by GadC at pH 9.0
with that accumulated at pH 5.0. What
fraction of the low pH activity is observed at
the higher pH?
a Glu-GABA antiporter
the GadC antiporter indeed functions only in acid environ-
ments, investigators compared its activity at a variety of pHs
with that of a different amino acid antiporter called AdiC,
which transports arginine out of cells under a broad
range of conditions. The results of monitoring transport for
10 minutes are presented in the graph.
Outside
Inside
cell
cell
3. Making Inferences
Would you say that the GadC
antiporter exhibits the same pH dependence as the
AdiC antiporter? If not, which antiporter is less
active at nonacid pHs?
GABA
4. Drawing Conclusions
Is the glutamate-GABA
antiporter GadC active at nonacid pHs?
5. Further Analysis
The GadC antiporter also
Glutamate
transports the amino acid glutamine (Gln). Do you
think this activity has any role to play in combating
low pH environments? How would you test this
hypothesis?
Chapter 5 Membranes 111
Substrate accumulation
(nmol per mg protein)
Inquiry & Analysis
Transcribed Image Text:How Hemorrhagic E. coli Resists the Acid Environment pH Sensitivity of the Glu-GABA Antiporter 300 GadC of the Stomach Adic 250- Recent years have been marked by a series of food poisoning outbreaks involving hemorrhagic (producing internal bleeding) strains of the bacterium Escherichia coli (E. coli). Bacteria are often a source of food poisoning, typically milder infections caused by food-borne strepto- coccal bacteria. Less able to bear the extremely acidic conditions encountered by food in the human stomach (pH 2), E. coli has not been as common a problem. The hemorrhagic strains of E. coli responsible for recent out- breaks seem to have evolved more elaborate acid-resistance 200 150 100 50 5 6 7 systems. How do hemorrhagic E. coli bacteria survive in the acid environment of the stomach? The problem they face, pH in essence, is that they are ions, many of which diffuse into their cells. To rid them- selves of these excess hydrogen ions, the E. coli cells use a clever system to pump hydrogen ions back out of their cells First, the hemorrhagic E. coli cells take up cellular hydrogen ions by using the enzyme glutamic acid decarbox- ylase (GAD) to convert the amino acid glutamate to Y-aminobutyric acid (GABA), a decarboxylation reaction that consumes a hydrogen ion. Second, the hemorrhagic E. coli export this GABA from their cell cytoplasm using called GadC (this transmembrane protein channel is called an antiporter because it transports two molecules across the membrane in opposite directions). However, to survive elsewhere in the human body, it is important that the Glu-GABA antiporter of hemorrhagic E. coli not function, lest it short-circuit metabolism. To see if submerged in a sea of hydrogen Analysis 1. Applying Concepts a. Variable. In the graph, what is the dependent variable? b. Substrate. What is a substrate? In this investigation, what are the substrates that are accumulating? c. pH. What is the difference in hydrogen ion concentration between pH 5 and pH 7? How many times more (or less) is that? Explain. 2. Interpreting Data a. Does the amount of amino acid transported in the 10-minute experimental interval (expressed as substrate accumulation) vary with pH for the arginine-transporting AdiC antiporter? For the glutamate-transporting GadC antiporter? b. Compare the amount of substrate accumulated by AdiC in 10 minutes at pH 9.0 with that accumulated at pH 5.0. What fraction of the low pH activity is observed at the higher pH? c. In a similar fashion, compare the amount of substrate accumulated by GadC at pH 9.0 with that accumulated at pH 5.0. What fraction of the low pH activity is observed at the higher pH? a Glu-GABA antiporter the GadC antiporter indeed functions only in acid environ- ments, investigators compared its activity at a variety of pHs with that of a different amino acid antiporter called AdiC, which transports arginine out of cells under a broad range of conditions. The results of monitoring transport for 10 minutes are presented in the graph. Outside Inside cell cell 3. Making Inferences Would you say that the GadC antiporter exhibits the same pH dependence as the AdiC antiporter? If not, which antiporter is less active at nonacid pHs? GABA 4. Drawing Conclusions Is the glutamate-GABA antiporter GadC active at nonacid pHs? 5. Further Analysis The GadC antiporter also Glutamate transports the amino acid glutamine (Gln). Do you think this activity has any role to play in combating low pH environments? How would you test this hypothesis? Chapter 5 Membranes 111 Substrate accumulation (nmol per mg protein) Inquiry & Analysis
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