Unfolding Individual Domains of BmrA, a Bacterial ABC Transporter Involved in Multidrug Resistance

Round 1

Reviewer 1 Report

The article is well written, I have only a small list of minor edits:

 

Line 22

NBD and TMD abbreviations are mentioned for the first time and should be explained (unscrambled) for the convenience of the reader.

Line 75

misspelling – said >> side

Line 80

misspelling – site >> side

Line 94

misspelling – transporter >> transporters

Line 136

it is more correct to use term “longer wavelengths”, rather than “higher wavelengths”

Line 144

the phrase “and the maximum red shifted by about 15 nm” does not seem correct. It should be rephrased. For example, “the absorption maximum shifted to the red region of the spectrum by 15 nm” or smth. similar.

Lines 172-173

“The anionic detergent SDS is able to form mixed micelles with the mild detergent DDM, eventually resulting in unfolding of soluble as well as TM domains.”

Here, an explanation is needed due to which the denaturation of the globular and transmembrane domains occurs, since the causes differ. Otherwise it looks like the globular domain is denaturing due to micelle formation, which is not the case.

Lines 232-234

Here the substitutions made and their effect on functional properties of the BmrA are described, however, it is not described by what criteria such mutations were chosen. This information is extremely important, since the choice of mutation is part of the approach proposed in this work to study domain denaturation in multidomain proteins. Moreover, these substitutions led to a change in the activity of the protein.

Author Response

see attached document

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments for author File: Comments.pdf

Author Response

see attached document

Author Response File: Author Response.pdf