On March 25th and 26th 1996, the fourth United KingdomAnalytical Ultracentrifuge Users’ Group Meeting was heldat the University of Leicester and hosted by the NationalCentre for Macromolecular Hydrodynamics. The focus areas of the meeting were twofold. One comprised new de-velopments in terms of instrumentation (the launch of thenew XL-I analytical ultracentrifuge) and analysis proce-dures (molecular mass algorithms and dilute solution con-formation algorithms). The other comprised specific ap-plications to biological macromolecules of both biomedi-cal and commercial importance. From this meeting a num-ber of articles were invited and the following collection ofrefereed papers is the result. These contributions testify tothe expansion of popularity and potential of the analyticalultracentrifuge in modern biomolecular science.

Introduction

Seven years on from an article by H. K. Schachman (1989)in which he announced the renaissance of the analyticalultracentrifuge, the technique has now been firmly re-es-tablished as a powerful tool for the analysis of biomolec-ular systems in terms of molecular mass, molecular massdistribution (polydispersity), mass action phenomena(interactions) and solution conformation. Since this time,a plethora of scientific papers has been published, togetherwith two books (Harding et al. 1992; Schuster and Laue1994).

To keep pace with these developments, in Europe, anannual Users’ Group meeting is held alternately in the

United Kingdom and the Federal Republic of Germany.The main focus of the United Kingdom symposia has beenin developments and applications to biomolecular systems,whereas the German symposia have considered also col-loidal and synthetic polymer systems. For several yearsspecial editions of Progress in Colloid and Polymer Sci-ence have been published based on the German symposia,and now for the first time a collection of invited papersbased on the meeting hosted by the United Kingdom is be-ing published in the European Biophysics Journal. The pa-pers are arranged in the order: instrumentation; analysis;and specific applications to biological macromolecules.

Developments in instrumentation

It was particularly pleasing to be able to use the 4th UKMeeting to host the European launch of the most recentmodern analytical ultracentrifuge – the Beckman OptimaXL-I, which has integrated Rayleigh interference and ab-sorption optics. This is described in the paper by Furst. Theaddition of the Rayleigh interference system permits moresensitive optical detection of concentration distributions inthe ultracentrifuge cell, and without the requirement of achromophore: the ability to make absorption and interfer-ence optical records should be particularly advantageousfor the analysis of interactions in mixed solute systems.For very concentrated systems – necessary, for examplefor the study of interacting systems of molecules where theaffinity is very low, the schlieren optical system, still avail-able only on older analytical ultracentrifuges, is the onlysolution. A modern data acquisition system for such an in-strument is the subject of the paper by Clewlow et al.

Developments in mass and conformation analysis

A series of papers describes developments in the analysisand interpretation of ultracentrifuge data. A paper by

Eur Biophys J (1997) 25: 305–306 © Springer-Verlag 1997

Olwyn Byron · Stephen E. Harding

New developments in analytical ultracentrifugation and related macromolecular modelling techniques

PREFACE

O. Byron (½)National Centre for Macromolecular Hydrodynamics, Department of Biochemistry, University of Leicester, University Road, Leicester LE1 7RH, UK

S. E. HardingNational Centre for Macromolecular Hydrodynamics, Department of Applied Biochemistry and Food Science, University of Nottingham, Sutton Bonington LE12 5RD, UK

Behlke provides a general overview followed by a paperby Behlke and Ristau which considers specifically the anal-ysis of interacting biopolymer systems. The analysis of mo-lecular mass is then considered, and Colfen and Hardingdescribe the use of two fundamental programmes,MSTAR-A and MSTAR-I, which handle absorption andinterference sedimentation equilibrium ASCII data that areproduced, for example, from the XL-A and XL-I ultracen-trifuge. These programmes are used to obtain weight, z-average and point average molecular weight information.The output data can then be easily transported to more spe-cific analysis programmes (self-association, polydispersedistribution etc.) where appropriate. Three further softwarepapers then follow. Harding et al. describe the ELLIPSsuite of PC algorithms for both simple ellipsoid of revolu-tion (ELLIPS1) and more advanced triaxial ellipsoid (ELLIPS2–4) representations of biomolecular conforma-tion in solution using both hydration dependent and inde-pendent “Universal” shape functions. For molecules andbiomolecular assemblies with more complicated shapes,modelling based on the hydrodynamic bead approximationis more appropriate, and Garcia de la Torre et al. describea PC routine SOLPRO which also uses hydration depen-dent and independent Universal shape functions. It is nowpossible to model molecular shape without the necessityalso to consider size and potential ambiguities therein. Another PC bead modelling algorithm BEAMS is also described by Spotorno et al. which uses the ancillary programmes PROMOLP and GRUMB for helping the user with both the choice of number and radius of beadsand in the interactive construction of spatially pre-definedmodels. ELLIPS, SOLPRO and BEAMS now provide acomplementary set of conformation algorithms for fairlyrigid biological macromolecules in solution. In the finalanalysis paper, Pavlov considers more general macro-molecular shapes as represented by polysaccharides, aclass of biomolecule with a huge range of possible struc-tural architectures, in terms of the concentration depen-dence of the sedimentation coefficient, and describes the importance of using the concentration dependence term ks (ml/g) in combination with the sedimentation co-efficient for the analysis of the equilibrium rigidity of ex-tra-rigid, rigid, semi-flexible and flexible chain poly-saccharides.

Applications to biomolecular systems

The largest section in this issue, happily, is devoted to re-ports on the successful use of the analytical ultracentrifugeand associated techniques in the study of biomolecularsystems, and we have attempted to order these from stud-ies on small interacting peptides, through to proteins, largeprotein assemblies, proteins and glycoproteins involved inthe immune response and finally the structure of large mu-cin glycoproteins.

Two papers on peptides are included. One, described byLiu et al. concerns an HIV-enhancer-binding peptide whichdimerises in the presence of a leucine zipper; the other, de-scribed by Thomas et al concerns a trimerising alpha-hel-ical coiled-coil phenylalanine zipper-type peptide. Colfenet al. then describe the heterodimeric properties of the elec-tron transferring flavoprotein (ETF), and the paper by Deanet al. describes how a point mutation (Asp165His), whichis distant from the subunit interface, changes the homo-hexameric wild-type glutamate dehydrogenase into a di-meric state. The theme of association and oligomeric struc-ture is continued with papers by Byron et al. on DT-dia-phorase, confirming the predominance of dimer and iden-tifying the presence of tetramer; Flood et al. on the self-association of α-actinin fragments; Aerts et al. on the ef-fect of increasing concentrations of amphiphilic and neu-tral detergent on the quaternary structure of the lens pro-tein α-crystallin; and Varley et al. on large molecularweight association products of the macrophage inflamma-tory protein-1α (MIP-1α) and a biologically active mutantthereof.

The proteins and glycoproteins involved in the immuneresponse are the subject of papers by Silkowski et al., whoconsider weak interactions between the cell adhesion mole-cules CD2 and CD48 and obtain results which complementthose from surface plasmon resonance, and Beavil andBeavil, who consider the conformation of a complex be-tween a human IgE fragment and the IgE high affinity re-ceptor Fcε RI-α. Keown et al. then consider the factorsunderpinning the stoichiometry of this interaction and ex-clude the possibility that steric hindrance by Cε2 accountsfor the unexpected 1:1 stoichiometry. The theme of gly-coproteins is considered by Jumel et al., who derive a lin-ear random coil model for the large colonic mucin glycop-roteins, based on sedimentation analysis in conjunctionwith size-exclusion chromatography coupled to laser lightscattering for molecular weight distribution analysis. Em-anating from the famous Uppsala laboratories of Svedbergand Tiselius, the techniques of ultracentrifugation and elec-trophoresis have been inextricably linked, particularly ascomplementary tools for the study of macro-ion diffusionphenomena, and the final article by Shepard et al. consid-ers very recent developments in this area.

Finally the editors of this special issue of the EuropeanBiophysical Journal are very grateful to the Managing Ed-itor, Dr. Peter Bayley, for his enthusiasm, help and patiencein bringing this collection of papers to press.

References

Harding SE, Rowe AJ, Horton JC (eds) (1992) Analytical ultracen-trifugation in biochemistry and polymer science. Royal Societyof Chemistry, Cambridge, UK

Schachman HK (1989) Analytical ultracentrifugation reborn. Nature341:259–260

Schuster T, Laue T (eds) (1994) Modern analytical ultracentrifuga-tion. Birkhauser, Boston, USA

306