<html><head></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; "><div><br></div><div>Hi Dave</div><div><br></div><div>To achieve a full thermodynamic characterization of protein stability is fairly arduous. But you could get a good idea of the relative stabilities of any mutants by following the temperature-induced unfolding of the proteins spectroscopically (e.g. using CD or Fluorescence). Comparison with the wild-type protein would give you a pretty decent idea of how much each mutation was costing you. Although this will not give you the complete thermodynamic information that DSC does (it's very difficult to estimate the heat capacity changes associated with protein unfolding using this kind of spectroscopic approach) it's fast, and much less heavy on material.</div><div><br></div><div>Best Regards,</div><div><br></div><div>Richard</div><div><br></div><div><span class="Apple-style-span" style="border-collapse: separate; color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: 2; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-border-horizontal-spacing: 0px; -webkit-border-vertical-spacing: 0px; -webkit-text-decorations-in-effect: none; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px; "><span class="Apple-style-span" style="border-collapse: separate; color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: 2; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-border-horizontal-spacing: 0px; -webkit-border-vertical-spacing: 0px; -webkit-text-decorations-in-effect: none; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px; "><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; "><div>Richard Kingston, PhD.</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">School of Biological Sciences</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">The University of Auckland</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">New Zealand</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">website: <a href="http://persephone.sbs.auckland.ac.nz/richard/lab/" style="color: blue; text-decoration: underline; ">http://persephone.sbs.auckland.ac.nz/richard/lab/</a></div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; "><br></div></div></span></span></div><br><div><div>On 4/06/2010, at 4:08 AM, Dj Scott wrote:</div><br class="Apple-interchange-newline"><blockquote type="cite"><o:smarttagtype namespaceuri="urn:schemas-microsoft-com:office:smarttags" name="country-region"><o:smarttagtype namespaceuri="urn:schemas-microsoft-com:office:smarttags" name="place"><div lang="EN-GB" link="blue" vlink="purple"><div class="Section1"><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Hello all in RASMB-land.<o:p></o:p></span></font></div><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">A question for you.<o:p></o:p></span></font></div><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; "><o:p> </o:p></span></font></div><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">If I make a void in a protein by mutating a larger residue to a smaller one in the hydrophobic core, what is the thermodynamic cost of this, and how would it be best to measure it.<o:p></o:p></span></font></div><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; "><o:p> </o:p></span></font></div><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">All thoughts welcome.<o:p></o:p></span></font></div><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Best<o:p></o:p></span></font></div><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; "><o:p> </o:p></span></font></div><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Dave.<o:p></o:p></span></font></div><p style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; "><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">>>>>>>>>>>>>>>>>>>>>>>>>>></span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Dr. David J. Scott</span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Associate Professor in Physical Biochemistry</span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">National Centre for Macromolecular Hydrodynamics</span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">School of Biosciences</span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">University of Nottingham</span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Sutton Bonington Campus</span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Leicestershire. LE12 5RD.</span></font> <br><st1:country-region w:st="on"><st1:place w:st="on"><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">United Kingdom</span></font></st1:place></st1:country-region><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">.</span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Phone: +44 (0) 115 951 6221</span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Fax: +44 (0) 115 951 6142</span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Email: <a href="mailto:dj.scott@nottingham.ac.uk" style="color: blue; text-decoration: underline; ">dj.scott@nottingham.ac.uk</a></span></font> <br><font size="2" face="Georgia"><span style="font-size: 10pt; font-family: Georgia; ">Web: <a href="http://www.nottingham.ac.uk/ncmh" style="color: blue; text-decoration: underline; ">www.nottingham.ac.uk/ncmh</a></span></font><o:p></o:p></p><div style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; margin-top: 0cm; margin-bottom: 0.0001pt; "><font size="3" face="Times New Roman"><span style="font-size: 12pt; "><o:p> </o:p></span></font></div></div><br><p style="margin-right: 0cm; margin-left: 0cm; font-size: 12pt; font-family: 'Times New Roman'; ">This message has been checked for viruses but the contents of an attachment may still contain software viruses which could damage your computer system: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation.</p></div><span><ATT00001..txt></span></o:smarttagtype></o:smarttagtype></blockquote><div><br></div></div><div><div>On 4/06/2010, at 4:08 AM, Dj Scott wrote:</div><br class="Apple-interchange-newline"><blockquote type="cite"><o:smarttagtype namespaceuri="urn:schemas-microsoft-com:office:smarttags" name="country-region">
<o:smarttagtype namespaceuri="urn:schemas-microsoft-com:office:smarttags" name="place">
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<div class="Section1"><p class="MsoNormal"><font size="2" face="Georgia"><span style="font-size:10.0pt;
font-family:Georgia">Hello all in RASMB-land.<o:p></o:p></span></font></p><p class="MsoNormal"><font size="2" face="Georgia"><span style="font-size:10.0pt;
font-family:Georgia">A question for you. <o:p></o:p></span></font></p><p class="MsoNormal"><font size="2" face="Georgia"><span style="font-size:10.0pt;
font-family:Georgia"><o:p> </o:p></span></font></p><p class="MsoNormal"><font size="2" face="Georgia"><span style="font-size:10.0pt;
font-family:Georgia">If I make a void in a protein by mutating a larger residue
to a smaller one in the hydrophobic core, what is the thermodynamic cost of
this, and how would it be best to measure it.<o:p></o:p></span></font></p><p class="MsoNormal"><font size="2" face="Georgia"><span style="font-size:10.0pt;
font-family:Georgia"><o:p> </o:p></span></font></p><p class="MsoNormal"><font size="2" face="Georgia"><span style="font-size:10.0pt;
font-family:Georgia">All thoughts welcome.<o:p></o:p></span></font></p><p class="MsoNormal"><font size="2" face="Georgia"><span style="font-size:10.0pt;
font-family:Georgia">Best<o:p></o:p></span></font></p><p class="MsoNormal"><font size="2" face="Georgia"><span style="font-size:10.0pt;
font-family:Georgia"><o:p> </o:p></span></font></p><p class="MsoNormal"><font size="2" face="Georgia"><span style="font-size:10.0pt;
font-family:Georgia">Dave.<o:p></o:p></span></font></p><p><font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">>>>>>>>>>>>>>>>>>>>>>>>>>></span></font>
<br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">Dr.
David J. Scott</span></font> <br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">Associate
Professor in Physical Biochemistry</span></font> <br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">National
Centre for Macromolecular Hydrodynamics</span></font> <br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">School
of Biosciences</span></font> <br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">University
of Nottingham</span></font> <br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">Sutton
Bonington Campus</span></font> <br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">Leicestershire.
LE12 5RD.</span></font> <br>
<st1:country-region w:st="on"><st1:place w:st="on"><font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">United Kingdom</span></font></st1:place></st1:country-region><font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">.</span></font>
<br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">Phone:
+44 (0) 115 951 6221</span></font> <br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">Fax:
+44 (0) 115 951 6142</span></font> <br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">Email:
<a href="mailto:dj.scott@nottingham.ac.uk">dj.scott@nottingham.ac.uk</a></span></font> <br>
<font size="2" face="Georgia"><span style="font-size:10.0pt;font-family:Georgia">Web:
<a href="http://www.nottingham.ac.uk/ncmh">www.nottingham.ac.uk/ncmh</a></span></font> <o:p></o:p></p><p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:
12.0pt"><o:p> </o:p></span></font></p>
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