Homework #3 KEY Systems Bioengineering I, Fall 2012, page 1 of 7

Homework 3: More gating and action-potential generation KEY

(100 points in total)
1. (80 points total) ION-CHANNEL GATING (Shaker K channels). Consider the whole-cell
current trace carried by Shaker K channels (with 4 m activation gates, and 1 h inactivation gate),
as shown below. The numerical data for this figure is given in a separate excel spreadsheet. The
I* relation for this cell is given by the equation: I* = 6 (pA/mV) * (V (-80 mV)), where V is
given in mV units. In the following subsections, you will carefully deduce the gating parameters
(m, m, h, h) at Vstep = -10 mV. Then, you will predict and plot the response of these channels
to an entirely new voltage protocol.

-10 mV

-100 mV

-100 mV

Homework #3 KEY Systems Bioengineering I, Fall 2013, page 2 of 7

1A. (20 pts) Using the I* relation, convert the trace into a time waveform for the probability of
the channel being open and plot it.

Obtained by dividing whole-cell current by I* at -10 mV between 0

and 8 msec.

1B. (20 pts) Fit a curve to the answer from Part 1A to calculate the timecourse of h (probability
that the h gate is enabled) for the time period between 0 and 8 msec. Plot the fit with a red line
and give the equation of your fitted curve (including all parameters). Interpret these parameters
to deduce the values of h and h at V = -10 mV. Assume that h = h(, -100 mV) = 1 for t < 0
ms.

Red curve = 0.0158 + (0.877 0.0158) * exp (-t / 2.403). Since h must
start from unity, h(t, -10 mV) = 0.018 + (1-0.018) * exp (-t / 2.403).
Hence, 1/2.403 ms-1 = h + h, and 0.018 = h / (h + h). Solving two
equations and two unknowns, h = 0.0075 ms-1 and h = 0.4087 ms-1.

Homework #3 KEY Systems Bioengineering I, Fall 2013, page 3 of 7

1C. (20 pts) Use the fit in part B to determine the time waveform for m (probability of m gates
being enabled) between 0 and 8 msec. Plot the waveform for m and deduce the values of
at -10 mV.

Since the overall probability waveform is given by m4 * h, in part B, the timecourse of m4 should
be obtained by dividing through the overall probability waveform by the timecourse of h as
determined in part B. This procedure yields the blue line above. To get m, we take the fourth
root of the blue line, yielding the thick gray line. This fits nicely to the function:
0.968 * (1 exp(-t * 2.3117), as shown by the thin red curve.
-1
Hence, 2.3117 ms = m + m, and 0.968 = m / (m + m). Solving two equations and two
unknowns, m = 2.2371 ms-1 and m = 0.0747 ms-1.

Homework #3 KEY Systems Bioengineering I, Fall 2013, page 4 of 7

1D. (20 pts) Having deduced the parameters above in parts A through C, see if you can predict
the behavior of the same cell for a different voltage protocol. Here, assume the channel has
reached steady state with respect to a holding potential of +100 mV. The valence for the m gate
is 4, and for the h gate is 2.5. Now give the explicit equation and plot for current between 0
and 8 ms, when the voltage is returned to -10 mV.

100 mV
-10 mV
-100 mV

Given the valences in question, steady-state values at 100 mV must be m = 1 and h = 0. Then,
the current during the 0-8 msec window would be a similar monoexponential function as in the
previous panels, except with different initial conditions.
m = 0.968 + (1 0.968) * exp(-t * 2.3117)
h = 0.018 + (0 0.018) * exp (-t * 0.4162)
I = I* m4 h, which is plotted in blue.

Homework #3 KEY Systems Bioengineering I, Fall 2013, page 5 of 7

2. (20 points total) ACTION-POTENTIAL GENESIS
In a certain type of smooth muscle in your gut, there are two main types of ion channels, Ca2+
channels and Ca2+-activated K channels. Your goal is to deduce the membrane potential of these
smooth muscles, because this potential determines the contraction strength of the smooth muscle.
A. (4 pts) Assume: (1) The Ca2+ channel is always fully open. (2) There are Nca = 50 Ca2+
channels per smooth muscle cell. (3) The battery-resistor single-channel conductance gca is 25e12 Siemens (amps/volt). (4) The channel is perfectly selective for Ca2+, with external and
internal Ca2+ concentrations held fixed at Co = 2e-3 and Ci = 1e-6, both in M units. Write the
equation (in variable form; dont plug in values yet!) for the whole cell Ca2+ current in a smooth
muscle cell.
Ica = Nca 1 gca (V (25/2) ln(Co / Ci))
B. (6 pts) The Ca2+-activated K channel has two identical and independent activation gates. Each
gate is NOT sensitive to voltage V, but rather to the simultaneous binding of two Ca2+ ions to its
intracellular loop. The on rate constant for two Ca2+ ions binding to the activation gate is given
by kon= 1e12 in units of transitions per (msec * M2). The off rate of Ca2+ from the activation gate
is given by koff = 1 in units of 1/msec. There are no inactivation gates. Assume: (1) There are
Nkca = 100 such channels per smooth muscle cell. (2) The battery-resistor single-channel
conductance gkca is 50e-12 siemens (amps/volt). (3) The channel is perfectly selective for K+,
with Nernst potential for K fixed at Vk = -50 mV. Write the equation (in variable form; dont
plug in values yet!) for the whole cell Ca2+-activated K+ current in a smooth muscle cell at steady
state. Assume the intracellular loop never binds 1 Ca2+ ion at a time (it either binds no Ca2+ ions
or 2 Ca2+ ions).
Ikca = Nkca gkca (Ci 2/ (Ci2 +koff / kon))2 (V Vk)

Homework #3 KEY Systems Bioengineering I, Fall 2013, page 6 of 7

C. (4 pts) Carefully plot the current-voltage relations from parts A and B (on the same figure)
from -100 mV to +100 mV. Then, deduce the voltage at which the smooth muscle will adopt a
steady state (the most positive stable fixed point), and indicate this voltage on your figure.
Recall that external and internal Ca2+ concentrations are held fixed at the values stated in part A.

Y-axis current in units of amperes. X-axis is transmembrane voltage V in mV. Red curve is kca
current, and blue curve is Ca2+ channel current. Presuming quasi-steady-state analysis may be
used, we sum the red and blue curves to yield the aggregate instantaneous I-V curve as the black
relation. We pick the stable steady-state voltage V at the intersection with the x-axis at V ~ 20
mV.

Homework #3 KEY Systems Bioengineering I, Fall 2013, page 7 of 7

D. (6 pts) Suppose the internal Ca2+ concentration (Ci) oscillates according to the function
in units of M, with t given in seconds (see graph below).
Derive the equation for V as a function of time, assuming validity of the quasi-instantaneous IV curve formalism of Noble, such that V resides at the moment-by-moment stable fixed point.
Plot the voltage waveform for the problem.

Solving for the voltage at which ICa + IK = 0 yields the equation

V = ( VCa NCa gCa + VK NK gK a(t) ) / (NCa gCa + NK gK a(t) ), where
a(t) = (Ci 2/ (Ci2 +koff / kon))2. This is plotted in black in the graph above.