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<TITLE>Re: [RASMB] upper concentration limit AUC</TITLE>
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Greeting, all<BR>
<BR>
Allen is indeed right - the analysis of concentrated solutions via SE analysis has indeed been pursued with significant success over the years, not least by Allen.<BR>
<BR>
I have even done a bit myself, introducing the INVEQ approach - i.e. fitting an inverted form of the basic SE equation to a <I>single data set,</I>, whereby values for Kd can be estimated even in the presence of non-ideality. See various studies:<BR>
<BR>
<FONT SIZE="2">Rensheng Luo, Beth Mann, William S. Lewis, Arthur Rowe, Richard Heath, Michael L. Stewart, Agnes E. Hamburger, Siva Sivakolundu, Eilyn Lacy, Pamela J. Bjorkman, Elaine Tuomanen, and Richard W. Kriwacki (2005) "Solution structure of choline binding protein A, the major adhesin of <I>Streptococcus pneumoniae</I>" EMBO Journal 24 1-10<BR>
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Arthur J Rowe (2005) "Weak Interactions: Optimal Algorithms for Their Study in the AUC" <I>in</I> "The Analytical Ultracentrifuge: Techniques & Methods" (eds, D J Scott, S E Harding & A J Rowe) pp 484-500. The Royal Society of Chemistry, London<BR>
<BR>
Christine Gee, Amanda Nourse, A-Yen Hsin, Qian Wu, Joel D Tyndall, Gary L Grunewald, Michael J McLeish, Jennfer L Martin "Disulphide-linked dimers of human adrenaline synthesising enzyme PNMT are catalytically active" Bikochim Biophys Acta (2005) 1750 82-92<BR>
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Zeng, W., Seward, H. E., Malnasi-Csizmadia, A., Wakelin, S., Woolley, R.J., Cheema, G. S., Basran, J., Patel, T. R., Rowe, A. J. and Bagshaw, C.R. (2006) Resonance Energy Transfer between Green Fluorescent Protein Variants: complexities revealed with Myosin Fusion Proteins. Biochemistry, 45, 10482-10491.<BR>
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Nyarko, Afua, Moshabi, Khédidja, Rowe, Arthur J., Leech, Andrew, Boter, Martin, Shirasu, Ken and Kleanthous, Colin (2007) TPR-Mediated Self-Association of Plant SGT1. Bikochemistry, 46 11331-11341<BR>
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Nyarko, Afua, Moshabi, Khédidja, Rowe, Arthur J., Leech, Andrew, Boter, Marta, Shirasu, Ken and Kleanthus, Colin (2007) TPR-Mediated Self-Association of Plant SGT1 Biochemistry 46 11331-11341<BR>
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Mullin, Nicholas P., Yates, Adam, Rowe, Arthur J., Mijmeijer, Bianca, Colby, Douglas, Barlow, Paul N., Walkinshaw, Malcolm D. and Chambers, Ian (2008) The pluripotency rheostat Nanog functions as a dimer. Biochem J <B>4</B> 227-231<BR>
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I will be reviewing the current 'state of the INVEQ art' at the Newcastle Meeting next week, hopefully for subsequent publication in Biophys Chem. New results include the estimation of both 2nd and 3rd virial coefficients, in addition to the Kd values for self-association. <BR>
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Regards to all<BR>
<BR>
Arthur<BR>
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Arthur J Rowe<BR>
Professor of Biomolecular Technology<BR>
NCMH Business Centre<BR>
University of Nottingham<BR>
School of Biosciences<BR>
Sutton Bonington<BR>
Leicestershire LE12 5RD UK<BR>
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Tel: +44 (0)115 951 6156<BR>
+44 (0)116 271 4502<BR>
Fax: +44 (0)115 951 6157<BR>
email: arthur.rowe@nottingham.ac.uk<BR>
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<BLOCKQUOTE><B>From: </B>Allen Minton <minton@helix.nih.gov><BR>
<B>Date: </B>Tue, 02 Sep 2008 09:00:58 -0400<BR>
<B>To: </B>Tom.Laue@unh.edu, "Beld, Joris" <beld@org.chem.ethz.ch><BR>
<B>Cc: </B>RASMB@server1.bbri.org<BR>
<B>Subject: </B>Re: [RASMB] upper concentration limit AUC<BR>
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<BLOCKQUOTE>Sedimentation equilibrium has been employed to study the properties<BR>
of proteins at high concentrations for many years. The following<BR>
publications are just for starters:<BR>
<BR>
R.C. Williams Jr. (1973) PNAS 70: 1506-1508. A very short<BR>
path-length cell was used to measure SE of hemoglobins A and S at<BR>
concentrations up to over 250 g/L.<BR>
<BR>
P.D. Ross et al. (1978) Biopolymers 17: 2285-8. Another type of<BR>
short path-length cell was used to measure SE of oxygenated and<BR>
deoxygenated hemoglobins A and S at different temperatures at<BR>
concentrations up to over 250 g/L.<BR>
<BR>
A.P. Minton & M. Lewis (1981) Biophys. Chem. 14 317-324. A<BR>
super-short path-length cell (ca .1 mm!) was used to measure SE of<BR>
hemoglobin and myoglobin at concentrations up to over 350 g/L.<BR>
<BR>
For interpretation of sedimentation equilibrium in highly nonideal<BR>
protein solutions (WAY beyond second virial coefficient), see Zorrilla et<BR>
al (2004). Sedimentation equilibrium in a solution containing an<BR>
arbitrary number of solute species at arbitrary concentrations: theory<BR>
and application to concentrated solutions of ribonuclease. Biophys. Chem.<BR>
108 89-100. <BR>
<BR>
Allen Minton<BR>
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