Supplement-2. Kinetic impact of [H+]o in H+ exporting pump with acceptor group Do

Reaction scheme, simplified from Fig. 1B.

Symbols:

S (1): So + Si;

SH (2): SoH + SiH;

DoH (3) and Do(4) as in Fig. 1B;

k1: transition probability from cytoplasmic H+ to SH, comprising several steps;

k2: transition probability from cytoplasmic SH to S; comprising several steps, e.g. initial photoisomerization (k2 = k20·light, k20: reference value) and transfer of H+ from SH to DoH;

k3: transition probability from DoH to Do, dissociation of H+ into external bulk solution;

k4: transition probability from Do to DoH, association of H+ from external bulk solution.

p1 p2, p3, p4: probabilities of indicated states to be occupied

Conditions and definitions:

p1 p2, p3, p4: probabilities of indicated states to be occupied

k3, k4 > k1, k2.

k4 = k40 [H+]o;

k40 = k4 at reference [H+]o;

K = k4/k3;

K0 = k40/k3;

p1 + p2 = 1;

p3 + p4 = 1.

Steady-state conditions:

0 = dp1/dt = -k1p1 + k2p2p4

0 = dp2/dt = k1p1 - k2p2p4;

0 = dp3/dt = k2p2p4 + k4p4 - k3p3;

0 = dp3/dt = -k2p2p4 - k4p4 + k3p3;

Turnover: I = k2p2p4.

After some algebra:

I = k1k2/(k1 + k1K + k2K);

Reciprocal:

1/I = (1+K)/k2 + K/k1.

At high pHo (K<1):

1/I = 1/k2 + K/k1,

= 1/k2 + K0[H+]o/k1;

DpHo induced change:

Parallel, straight lines with slope 1/(k20·light) and vertical displacements K0[H+]o/k1 from origin (Fig. 2B),

q.e.d.

1