Rate equations for the gross influx of -aminoisobutyric acid (AIB) into mouse cerebrum slices containing AIB have a first-order term for unsaturable concentrative influx, identical to the corresponding term for unloaded slices, and a modified Michaelis-Menten term,
Vmax/(1+
K
t
/
S), for saturable concentrative influx. [
Vmax v
L
(1+
K
t
/
S), where
v
L
=saturable component of influx,
S=AIB concentration in medium, and
K
t
=Michaelis constant for unloaded slices.] Below a tissue AIB (
T) of 19 µmol/g final wet weight,
Vmax increases linearly following
Vmax=
V
1+
m
1
T; above that value,
V
max is virtually constant. The transition is sharp. This equation is consistent with a carrier model for active transport. At the transition, intracellular AIB is about 1 molecule for every 70 amino acid residues of tissue protein, vastly more than could be accommodated by AIB-binding sites in cell membranes. The transition may come from a slow process that does not fill all sites when the tissue AIB is below the transition concentration, or from an AIB-induced phase transition in the membrane.Nomenclature AIB
-aminoisobutyric acid
-
A
radioactivity of reference; unspecified amino acid
-
C
counts in tissue sample; carrier for transport
-
C
i
carrier in form that reacts with intracellular substrate
-
C
o
carrier in form that reacts with extracellular substrate
-
C
R
counts in reference
-
CS
complex of substrate with carrier
- (
CS)
i
complex of substrate with carrier in form
C
i
- (
CS)
o
complex of substrate with carrier in form
C
o
-
G
counts per gram of tissue
- HEPES
N-2-Hydroxyethylpiperazine-
N-2-ethanesulfonic acid
-
k
u
rate constant for first-order unsaturable uptake
-
K,
K
,
K
,
K
,
K
d
adjustable parameters in Eqs. (9)–(13) for
v, analogous to the Michaelis constant
-
K
d
dissociation constant
-
K
t
Michaelis constant for saturable uptake
-
K
t
Michaelis constant for gross saturable uptake by tissue containing substrate
-
m
1,
m
2
slope in Eq. (5) or (6) expressing dependence of
Vmax on
T or
T
i
w
in Region 1 or 2
-
M
binding site for amino acid A
-
n
number of data points
-
P
number of parameters to be determined; parameter in Stein's (1981) equation, Eq. (17) in this paper
-
P
1,
P
2,
P
12
property of tissue with unoccupied binding sites, property of tissue with occupied binding sites, property of tissue with both unoccupied and occupied binding sites, respectively
-
Q
parameter in Stein's (1981) equation, Eq. (17) in this paper
-
r
Pearson's correlation coefficient
- Relative error RE
=100{[(observed quantity – calculated quantity)/calculated quantity]
2/(
n –
P)}
1/2
-
S
concentration of substrate in medium; transport substrate
-
S
i
intracellular transport substrate
-
S
int
AIB in medium corresponding to intracellular AIB at intersection
-
S
o
extracellular transport substrate
-
T
observed concentration of substrate in tissue including substrate in extracellular space and adherent fluids
-
T
i
intracellular concentration of substrate
-
T
int
tissue AIB corresponding to intracellular AIB at intersection
-
T
i
w
,
T
i
/30
intracellular concentration of substrate with
w% (30%) extracellular and adherent fluids
-
U
observed uptake of labeled substrate by incubated tissue including substrate in extracellular and adherent fluids
-
U
R
observed uptake of labeled substrate referred to concentration of substrate in medium
-
U
max
adjustable parameter in Eqs. (9)–(15) for
v, analogous to the Michaelis-Menten maximum rate,
V
max
-
v
influx of substrate
-
v
L
gross influx of substrate into tissue containing substrate
-
v
L
contribution of saturable component to gross influx into tissue containing substrate
-
v
incremental influx, that is, gross influx into tissue that contains substrate minus influx under the same conditions into tissue that does not contain substrate
-
V
1,
V
2
intercept in Eq. (5) or (6) expressing dependence of
Vmax on
T or
T
i
w
in Region 1 or 2, respectively
-
V
max
maximum rate in Michaelis-Menten equation
-
Vmax
apparent maximum rate defined by
Vmaxvmax(1+
K
t
/
S)
-
Vmax 1,
Vmax 2
apparent maximum rate in Region 1 or 2, respectively
-
Vint
apparent maximum rate at intersection defining boundary between Regions 1 and 2
-
w
weight of incubated tissue
-
W
d
dry weight of tissue expressed as fraction by weight
-
W
e
extracellular and surface space of incubated tissue expressed as percent by weight
-
, ,
adjustable parameters in modified expressions for gross unsaturable influx into tissue containing substrate
-
, , ,
exponents of
S or
T in Eqs. (9)–(13) for
v
-
parameter in Stein's (1981) equation, Eq. (17), corresponding more or less to
m
1
For my wife, Lynn.
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