Hi!
Thanks to Dr. Herman I realized that I unintentionally made an overstatement in my "questions after
symposium" letter. I believe it’s important to clarify the issue. Dr. Herman kindly allowed me to post the
text of our communications. Here it is, without any modifications:
>Dear Dr. Kandel
>>I read with great interest your questions that came up during the preparation
>of the meeting summary for the recent GFP meeting in New Jersey. One of your
>comments and answers intrigued me. You asked whether the ability of the GFP
>tag to domerize leads to artifacts, and you site as an example the fact that
>GFP will make Bcl-2 dimerize with itself or Bax at the expense of other
>natural counterparts (e.g. Bad or BclX). I was wondering if you could tell me
>the source of this finding. Is this informatino published somewhere, or are
>these experiments you have done personally? We have submitted a paper showing
>specific interaction between Bcl-2 and Bax in mitochondira of cingle living
>cells, so I am intersted in the origin of this statement.
>>I would appreciate it of you could respond to me at my office email-
>bhgf at med.unc.edu.>>Sincerely yours,
>>Brian Herman, Ph.D.
>Professor
>Department of Cell Biology & Anatomy
>CB # 7090, 232 Taylor Hall
>University of North Caroilina
>Chapel Hill, NC 27599
>919-962-0350 (voice)
>919-966-1856 (FAX)
>bhgf at med.unc.edu (email)
>
Dear Dr. Herman,
In the summary of questions that I had remaining after the GFP
meeting I tried to mention the artifacts that to the best of my
understanding could (but not necessarily have to) occur. After I read you
letter, I realized that instead of writing " extra dimerization domain (=
GFP tag) COULD make bcl2 dimerize preferentially with another bcl2 or bax
molecule that contains such a tag" I used "extra dimerization domain (= GFP
tag) CAN make bcl2 dimerize preferentially with another bcl2 or bax molecule
that contains such a tag". I apologize if in my original statement I sounded
like I have a definitive knowledge that this artifact exists.
My original intent was to express concern that a clear possibility
for this artifact exists (it seems quite possible that proteins bound by two
domains might be bound more tightly than those bound by only one) ,and I AM
NOT AWARE (maybe it's my personal impression) of this possibility being
properly ruled out in the studies presented at the Symposium. I would like
to thank you for pointing out my overstatement. I realize that it could have
confused others as well, and a clarification of this matter may be
important. I would like to ask your permission to post your letter and my
reply to the same newsgroup where my original statement appeared. If you
prefer, I could post only my clarification with a reference to you, as the
person who pointed out my mistake. In any case, I would like to post the
clarification as soon as possible.
I understand that presentation at the Symposium does not include all
the data accumulated in the field, and the possibility of the above
mentioned artifacts could have been ruled out by direct experiments. In this
case, I would like to ask you for the permission to mention such experiments
in my clarification letter: I believe this could set up an example for
future studies in similar systems. Thank you for your help.
Sincerely,
Eugene Kandel.
>Dear Dr Kandel
>>You may post my email and your response.
>>Brian Herman
