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<DIV><SPAN class=020583415-16092002><SPAN class=990025517-16092002></SPAN>T<SPAN
class=990025517-16092002>o </SPAN>John<SPAN class=560111818-17092002>,</SPAN>
<SPAN class=990025517-16092002><SPAN class=560111818-17092002>all </SPAN>DCDT+
users<SPAN class=560111818-17092002> <SPAN class=820054219-19092002>and
</SPAN>all<SPAN class=990025517-16092002> </SPAN>interested colleagues<SPAN
class=560111818-17092002>, </SPAN></SPAN></SPAN></SPAN></DIV>
<DIV><SPAN class=020583415-16092002><SPAN class=990025517-16092002><SPAN
class=560111818-17092002><SPAN class=560111818-17092002></SPAN> </DIV>
<DIV><SPAN class=020583415-16092002><SPAN
class=560111818-17092002></SPAN></SPAN> </SPAN></SPAN></SPAN><SPAN
class=020583415-16092002>John, I would like to have your comment in the
following three experiments using RNAse<SPAN class=560111818-17092002>A
</SPAN>as a model and DCDT+ <SPAN class=990025517-16092002>as the method
</SPAN>to do data analysis.</SPAN></DIV>
<DIV><SPAN class=020583415-16092002></SPAN> </DIV>
<DIV><SPAN class=020583415-16092002>Experiment 1: <SPAN
class=560111818-17092002>The p</SPAN>urpose is to check the stability of
RNAse<SPAN class=560111818-17092002>A</SPAN><SPAN class=560111818-17092002>. The
comparison is made between t</SPAN>he protein <SPAN
class=560111818-17092002><SPAN class=820054219-19092002>left</SPAN> </SPAN>at
room temperature for <SPAN class=990025517-16092002>a
</SPAN>couple <SPAN class=820054219-19092002>of </SPAN>days and <SPAN
class=560111818-17092002>that </SPAN>stor<SPAN
class=820054219-19092002>age</SPAN> in the <SPAN
class=820054219-19092002>freezer.</SPAN></SPAN></DIV>
<DIV><SPAN class=020583415-16092002><SPAN class=020583415-16092002>Experiment
2: <SPAN class=990025517-16092002>Question: </SPAN><SPAN
class=990025517-16092002>Can <SPAN class=560111818-17092002>the
</SPAN>denatur<SPAN class=560111818-17092002>ed</SPAN></SPAN><SPAN
class=990025517-16092002> </SPAN>RNase<SPAN class=560111818-17092002>A<SPAN
class=820054219-19092002> </SPAN></SPAN>(has 4 s-s bond) in GuHCl <SPAN
class=990025517-16092002>be refolded back </SPAN>to its native state in the
absence <SPAN class=820054219-19092002>of </SPAN>reducing agent<SPAN
class=560111818-17092002>?</SPAN></SPAN></SPAN></DIV>
<DIV><SPAN class=020583415-16092002><SPAN class=020583415-16092002><SPAN
class=020583415-16092002>Experiment 3: <SPAN
class=990025517-16092002><SPAN class=990025517-16092002>Question: </SPAN><SPAN
class=990025517-16092002>Can <SPAN class=560111818-17092002>the
</SPAN></SPAN>denatur<SPAN class=560111818-17092002>ed</SPAN></SPAN><SPAN
class=990025517-16092002> </SPAN>RNase<SPAN class=560111818-17092002>A<SPAN
class=820054219-19092002> </SPAN></SPAN>(has 4 s-s bond) in GuHCl <SPAN
class=990025517-16092002>be refolded back </SPAN>to its native state in the
presence of reducing agent <SPAN
style="FONT-FAMILY: 'Times New Roman'; FONT-SIZE: 12pt; mso-fareast-font-family: 'Times New Roman'; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA">TBP
(Tri-n-butylphosphine)<SPAN
class=560111818-17092002>.</SPAN></SPAN></SPAN></SPAN></SPAN></DIV>
<DIV><SPAN class=020583415-16092002><SPAN class=020583415-16092002><SPAN
class=020583415-16092002><SPAN
style="FONT-FAMILY: 'Times New Roman'; FONT-SIZE: 12pt; mso-fareast-font-family: 'Times New Roman'; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA"><SPAN
class=560111818-17092002></SPAN></SPAN></SPAN></SPAN></SPAN> </DIV>
<DIV><SPAN class=990025517-16092002>The major issue here is how <SPAN
class=560111818-17092002>I </SPAN>can differentia<SPAN
class=560111818-17092002>te</SPAN> the<SPAN class=560111818-17092002> <B
style="mso-bidi-font-weight: normal"><I
style="mso-bidi-font-style: normal"><SPAN
style="FONT-FAMILY: Symbol; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol"><SPAN
style="mso-char-type: symbol; mso-symbol-font-family: Symbol">D</SPAN></SPAN>S<?xml:namespace
prefix = o ns = "urn:schemas-microsoft-com:office:office"
/><o:p></o:p></I></B></SPAN> difference due to scan cho<SPAN
class=560111818-17092002>sen</SPAN> <SPAN
class=560111818-17092002>from</SPAN> <SPAN
class=560111818-17092002>the <B style="mso-bidi-font-weight: normal"><I
style="mso-bidi-font-style: normal"><SPAN
style="FONT-FAMILY: Symbol; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol"><SPAN
style="mso-char-type: symbol; mso-symbol-font-family: Symbol">D</SPAN></SPAN>S<o:p></o:p></I></B><B
style="mso-bidi-font-weight: normal"><I
style="mso-bidi-font-style: normal"><SPAN
style="FONT-FAMILY: Symbol; FONT-SIZE: 14pt; mso-fareast-font-family: 'Times New Roman'; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol; mso-bidi-font-size: 12.0pt; mso-bidi-font-family: 'Times New Roman'"><SPAN
style="mso-char-type: symbol; mso-symbol-font-family: Symbol">
</SPAN></SPAN></I></B>difference due to </SPAN><SPAN
class=560111818-17092002>real</SPAN> experimental finding<SPAN
class=820054219-19092002> (between the control and the
sample)</SPAN>.</SPAN></DIV>
<DIV><SPAN class=990025517-16092002></SPAN> </DIV>
<DIV><SPAN class=020583415-16092002><SPAN class=990025517-16092002></SPAN><FONT
size=2>I<SPAN class=990025517-16092002> am please<SPAN
class=560111818-17092002>d</SPAN> to say that</SPAN><SPAN
class=990025517-16092002>, using the criteria that I have stated<SPAN
class=820054219-19092002>,</SPAN><SPAN class=500034417-17092002><SPAN
class=560111818-17092002> e.g. m</SPAN>aximized or magnified the<SPAN
style="FONT-FAMILY: Symbol; FONT-SIZE: 14pt; mso-fareast-font-family: 'Times New Roman'; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol; mso-bidi-font-size: 12.0pt; mso-bidi-font-family: 'Times New Roman'"><SPAN
style="mso-char-type: symbol; mso-symbol-font-family: Symbol"><STRONG><EM>
</EM></STRONG><FONT size=2><FONT face=Arial><B
style="mso-bidi-font-weight: normal"><I
style="mso-bidi-font-style: normal"><SPAN
style="FONT-FAMILY: Symbol; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol"><SPAN
style="mso-char-type: symbol; mso-symbol-font-family: Symbol">D</SPAN></SPAN>S<SPAN
class=560111818-17092002> </SPAN></I></B></FONT><FONT face=Arial>difference due
to scan cho<SPAN
class=560111818-17092002>sen</SPAN> </FONT></FONT></SPAN></SPAN>(<SPAN
class=820054219-19092002>use</SPAN> a small<SPAN
class=560111818-17092002>er</SPAN> number for denominator<SPAN
class=560111818-17092002> when convert<SPAN
class=820054219-19092002>ed</SPAN></SPAN><SPAN class=560111818-17092002> to
%</SPAN>)<SPAN class=560111818-17092002>,</SPAN> <SPAN
style="FONT-FAMILY: 'Times New Roman'; FONT-SIZE: 12pt; mso-fareast-font-family: 'Times New Roman'; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA"><SPAN
class=560111818-17092002><FONT face=Arial size=2>m</FONT></SPAN>inimized or
play down the <FONT face="Times New Roman"><FONT size=2><B
style="mso-bidi-font-weight: normal"><I
style="mso-bidi-font-style: normal"><SPAN
style="FONT-FAMILY: Symbol; mso-ascii-font-family: 'Times New Roman'; mso-hansi-font-family: 'Times New Roman'; mso-char-type: symbol; mso-symbol-font-family: Symbol"><SPAN
style="mso-char-type: symbol; mso-symbol-font-family: Symbol">D</SPAN></SPAN>S<SPAN
class=560111818-17092002> </SPAN></I></B></FONT>difference<SPAN
class=560111818-17092002> between <SPAN class=820054219-19092002>the
</SPAN>control and the sample</SPAN> (<SPAN class=820054219-19092002>use</SPAN>
a large<SPAN class=560111818-17092002>r</SPAN> number for denominator<SPAN
class=560111818-17092002> when convert<SPAN class=820054219-19092002>ed</SPAN>
to </SPAN><SPAN
class=560111818-17092002>%</SPAN> </FONT></SPAN></SPAN><FONT
face="Times New Roman" size=3><SPAN
class=500034417-17092002>)</SPAN></FONT></SPAN><SPAN
class=560111818-17092002><FONT face="Times New Roman" size=3>,</FONT></SPAN> the
results are <SPAN class=820054219-19092002>pretty much </SPAN><SPAN
class=820054219-19092002>I have</SPAN> expected. Do you think I <SPAN
class=990025517-16092002>try to push </SPAN>the instrument limit to<SPAN
class=990025517-16092002>o far to</SPAN> <SPAN
class=990025517-16092002>obtain</SPAN> these results by manipulat<SPAN
class=820054219-19092002>ing</SPAN> the <SPAN
class=990025517-16092002>treatment of data</SPAN>? </FONT></SPAN></DIV>
<DIV><SPAN class=020583415-16092002> </DIV>
<DIV><SPAN class=560111818-17092002>s20,w values reported here are the results
of fitting dcdt.</SPAN></DIV>
<DIV><SPAN class=560111818-17092002></SPAN> </DIV>
<DIV><SPAN class=020583415-16092002>Thank you in advance<SPAN
class=560111818-17092002> for your comment or advice.</SPAN></SPAN></DIV>
<DIV><SPAN class=020583415-16092002></SPAN> </DIV>
<DIV><SPAN class=020583415-16092002><SPAN
class=020583415-16092002>Chris</SPAN></SPAN></DIV>
<DIV><SPAN class=020583415-16092002></SPAN>
<P>--------------------------------------------------------------------------
<BR>Christopher Chin <BR>Manager, XLA-Analytical Ultracentrifugation facility
<BR>Sealy Center for Structural Biology <BR>HBC&G, 5.134 MRB.UTMB,
Galveston,T<SPAN class=560111818-17092002>X</SPAN> 77555-1055
<BR>cchin@utmb.edu, 409-772-1693, efax 708-585-1920
<BR>---------------------------------------------------------------------------
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