Does the membrane potential usually quoted for Gram negative bacteria (e.g., E. coli) refer to the potential across both membranes? - If yes, then does the potential fall more over the inner or outer membrane? Are there any studies that estimate/measure the relative fraction for each membrane?

I assume that the measurements of potential are usually carried out by inserting an electrode inside the cell and measuring the difference with the outside, this would make the answer to the first question yes.

  • 1
    $\begingroup$ There is probably a clue in the context. Generally though potential refers to the intra-extra cellular potential (i.e in this case both membranes). Is there a particular source you can cite that is ambiguous? $\endgroup$
    – James
    Apr 27, 2015 at 10:33
  • $\begingroup$ This is a good answer to the first part of the question. My motivation for asking is partly that I would like to understand the membrane potential experienced by a trans-membrane protein present only on the inner membrane. Even a rough educated guess (with reasoned argument) would be helpful. For example: Would the presence more proteins that are driven by membrane potentials being located on only one of the membranes be a good indicator that the the potential would be likely to fall mostly over that membrane? If no literature exists, is it reasonable to construct an argument in this way? $\endgroup$
    – someone
    Apr 27, 2015 at 16:06
  • $\begingroup$ I suggest you edit this question to ask only about Gram-staining and open a new question that asks about Gram-negative membrane potential. $\endgroup$
    – James
    May 5, 2016 at 1:23

2 Answers 2


Short answer
The distinction between Gram positive (Gram+) and negative bacteria (Gram-) has absolutely nothing to do with membrane potentials; it is all about the Gram staining procedure.

The Gram staining was named after the Danish bacteriologist Hans Christian Gram, who originally devised it in 1882 (Gram, 1884). Gram staining is a common technique used to differentiate two large groups of bacteria based on their different cell wall constituents. The Gram stain procedure distinguishes between Gram+ and Gram- groups by coloring these cells violet or red, respectively (Fig. 1).

Gram+ bacteria stain violet due to the presence of a thick layer of peptidoglycan in their cell wall, which retains the crystal violet these cells are stained with. Gram-, on the other hand stain red, which is attributed to a thinner peptidoglycan wall, which does not retain the crystal violet during the decoloring process.

Gram staining procedure
The procedure involves staining the cells with a crystal violet dye. Next, an iodine solution (iodine and potassium iodide) is added to form a complex between the crystal violet and iodine, which is insoluble in water. Then, a decolorizer, such as ethyl alcohol, is added to the sample, which dehydrates the peptidoglycan layer, shrinking and tightening it. The crystal violet-iodine complex is not able to penetrate this tightened peptidoglycan layer, and is thus trapped in the cell in Gram+ bacteria. In Gram- bacteria, however, the thinner peptidoglycan layer does not retain the crystal violet-iodine complex and the color is lost. Lastly, a counterstain, such as the weakly water soluble safranin, is added to the sample, staining it red. Since the safranin is lighter of color than crystal violet, it does not disrupt the purple coloration in Gram+ cells, but it does stain the decolorized Gram- cells red.

Gram + and -
Fig. 1. Gram+ bacteria (left) and Gram- on the right. source: Lab Tests Online

- Gram, Fortschritte der Medizin (1884)

  • $\begingroup$ Good answer to the question. However it's a classic XY problem. OP is actually looking for the membrane potential for the periplasmic space to the cytosol. $\endgroup$
    – James
    May 5, 2016 at 1:22
  • $\begingroup$ @James Thanks! Regarding your point, I agree and that's why I upvoted the other answer here. $\endgroup$
    – AliceD
    May 5, 2016 at 7:37

The following is not an answer to the original question: "Are Gram negative bacteria classified as such because of their negative membrane potential?" but to the questions later in the text.

Usually the membrane potential is given for the inner cytosolic part and the extracellular space, for E. Coli it is around -120 mV; see also this article.

Due to the small size of E. Coli it is very difficult to reliably and accurately measure the membrane potential.

For the periplasm only the Donnan potential is reported, it ranges between 5 and 100 mV, typically around 25 mV.

I don't know about any report measuring the relative contribution but I would guess that the inner part makes up most of the total potential.


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