WASHINGTON UNIVERSITY

TU HENRY SHAW SCHOOL
     OF BOTANY

SAINT LOUIS

March 15, 1955

Dr, Rosalind E, Franklin
Birkbeck College
University of London
21 Torrington Square, W,C.l,
London
England

Dear Dr, Franklin:

deal of interest,
a great deal of what we want to know about the relations between TMV
and the nonvirus proteins.

I have gone over the material which you send me with a great

Your investigations certainly promise to tell us

  Although there are a number of extremely interesting points which
I shall want to write you about, this letter will be restricted to the
immediate issue of the paper which you have prepared on your results
with B8.  After reading your manuscript it appears evident that a
good deal of confusion now exists with regard to the abnormal proteins
related to TMV. The facts as I see them are these,

1)

W-infected plants produce at least two and'probably three

low-molecular weight abnormal nonvirus proteins which are immuno-
chemically related to TMV and which polymerize to form "V-like rods.

     As noted in my revision of your manuscript, there is agreement
on the existence of at least two of the above proteins between myself,
Jeener and Takahashits ori inal publication (her, Jour. of Botany, 40,
85) , %he correlations + etween my terminology and Takahashits, with
respect to the two proteins, is presented in my revision of your
manuscript,

2)

3)

    Very recently the paper by Delwiche, Takahashi, Newmark, et al,
(Biochim. Biophys. Acta l6, 127) claims that only a single abnormar -
protein can be found, However, they offer no data to substantiate this
claim and the method of preparation which they use is identical with
one of the methods which we have employed to isolate two separate non-
virus oomponents (B3 and B6),
been concerned over this discrepancy and have examined the behavior of
mixtures of B3 and B6.
a single high-molecular weight component which migrates in a homogeneous
fashion in the Pbelius apparatus at pH 7.0,
components will form a co-polymer,
made by first preparing a homogeneous preparation of B3  and then
polymerizing it to form B8,
polymerization is induced it is likely that a complex co-polymer will
be formed which is difficult to distinguish from B8.

    Since seeing the Delwiche paper I have

We find that such mixtures polymerize to form

In other words, the two

   Our own preparations of B8 are
Unless B3 and B6 are separated before

I suspect very

-2- March 15, 1955

strongly that the protein described by Delwiche et al,  and which was
used by Rich in his work vas very likely such a =-polymer containing
both B3 and B6 in an indefinite ratio,

   4)  For these reasons the Babnormal proteinn of Rich and Delwiche
should not be considered the equivalent of B8.
workers, including Takahashi, are not aware of the occurrence of B6
and its participation in the formation of a co-polymer, it is entirely
likely that successive preparations from their laboratories may
contain variable ratios of B3 and B6,

equivalent to our B8,
extracts are manipulated and it is possible to lose most of the B6 in
a preparation as a result of lengthy processing,
that the major variation in the preparations produced by the other
laboratories would be a fluctuation in the relative amount of B6 present
in the polymerized "abnormal proteinw and that in some cases the B6
content may be zero without their being aware of it,

your manuscript,
that you will feel free to condense it as far as possible,
also made a few minor changes in the first page or two of your
manuscript to avoid statements which cannot be supported by our own
data, This includes Richst assertion that the chemical composition of
the abnormal protein and TMV are identical,
statement in view of the likelihood that their protein is not of
constant composition,

Furthermore, if these

Thus, the reparation which

Takahashi has sent you may contain little or no B z and is, therefore,

We find that B6 is readily denatured when

Hence, I would expect

1 have tried to incorporate some of these ideas in an addition to
      What I have added is probably unduly long and I hope

I have

This is a very doubtful

Because of the material which I have added to the manuscript I

think that it would be wise if my name were added to the paper.  !'he
results are extremely interesting and I am happy for the opportunity
to be associated with the publication,
  Also included herewith are galley proofs of two papers on the
nonvirus proteins which will appear in the March 20th issue of the
JOURNAL OF GENERAL PHYSIOLOGY.
can have     idea of the evidence which we now have on the wU%&&?- rhd
and properties of the nonvirus proteins,

I am sending you these now so that you

some

  Under separate cover I am sending you about 10 milligrams more
of the B8 preparation which you now have,  After some trials I decided
that the best orientated preparation appeared to be a pellet of the
protein spun down in the ultracentrifuge.  The material is being sent
to you in this form,
in particle alignment by forcing this material into a capillary,
any event, the pellet which I am sending you is the most concentrated
form in which I can prepare B8 and should make your manipulations
easier to carry out.

I hope that you will be able to get an improvement

In

-+d-L-

-3 - March 15, 1955

   We are very much interested in obtaining good amino acid analyses
on this preparation of B8,
try to salvage as much material as you can from your operations and
return it to me when it is no longer useful to you.  Regardless of
the physical state of the preparation we can use it for amino acid
anal ysi 8 ,

Consequently,  I should like to ask you to

   I hope you will write again soon and let me know what you think
of the situation, In a little while I shall write you concerning
some of the ideas which I have on the possible relations between the
abnormal polymerized proteins and TMV nucleoprotein, Meanwhile, 1
shall look forward with interest to learning of your latest results.

Sincere yours,

3-cw.

Barry Commoner

BC: bl
Encl ,