[RASMB] upper concentration limit AUC

[Arthur Rowe]

Greeting, all

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.

I have even done a bit myself, introducing the INVEQ approach - i.e. fitting
an inverted form of the basic SE equation to a single data set,, whereby
values for Kd can be estimated even in the presence of non-ideality. See
various studies:

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 Streptococcus
pneumoniae" EMBO Journal 24 1-10

Arthur J Rowe (2005) "Weak Interactions: Optimal Algorithms for Their Study
in the AUC" in "The Analytical Ultracentrifuge: Techniques & Methods" (eds,
D J Scott, S E Harding & A J Rowe) pp 484-500. The Royal Society of
Chemistry, London

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

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.

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

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

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 4 227-231

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.

Regards to all

Arthur


--
*******************************************************
Arthur J Rowe
Professor of Biomolecular Technology
NCMH Business Centre
University of Nottingham
School of Biosciences
Sutton Bonington
Leicestershire LE12 5RD   UK

Tel:        +44 (0)115 951 6156
          +44 (0)116 271 4502
Fax:        +44 (0)115 951 6157
email:      arthur.rowe at nottingham.ac.uk
Web:        www.nottingham.ac.uk/ncmh/business
*******************************************************


From: Allen Minton <minton at helix.nih.gov>
Date: Tue, 02 Sep 2008 09:00:58 -0400
To: Tom.Laue at unh.edu, "Beld, Joris" <beld at org.chem.ethz.ch>
Cc: RASMB at server1.bbri.org
Subject: Re: [RASMB] upper concentration limit AUC


Sedimentation equilibrium has been employed to study the properties
of proteins at high concentrations for many years.  The following
publications are just for starters:

R.C. Williams Jr. (1973) PNAS 70: 1506-1508.  A very short
path-length cell was used to measure SE of hemoglobins A and S at
concentrations up to over 250 g/L.

P.D. Ross et al. (1978) Biopolymers 17: 2285-8.  Another type of
short path-length cell was used to measure SE of oxygenated and
deoxygenated hemoglobins A and S at different temperatures at
concentrations up to over 250 g/L.

A.P. Minton & M. Lewis (1981) Biophys. Chem. 14 317-324. A
super-short path-length cell (ca .1 mm!) was used to measure SE of
hemoglobin and myoglobin at concentrations up to over 350 g/L.

For interpretation of sedimentation equilibrium in highly nonideal
protein solutions (WAY beyond second virial coefficient), see Zorrilla et
al (2004).  Sedimentation equilibrium in a solution containing an
arbitrary number of solute species at arbitrary concentrations: theory
and application to concentrated solutions of ribonuclease. Biophys. Chem.
108 89-100.  

Allen Minton





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