Michel Laurent

Electrochemical Determination of pKa of N-Bases in Ionic Liquid Media

Two electrochemical techniques have been used to measure the pK(a) of N-bases in several ionic liquids (ILs). The first method corresponds to a potentiometric titration of a strong acid with the N-base using a platinized Pt indicator electrode immersed in the IL solution and maintained under dihydrogen atmosphere via gas bubbling. The second approach involves performing cyclic voltammetry at a platinized Pt electrode in a solution containing both strong acid and the conjugate weak acid of the N-base. Values of pK(a) obtained by one or the other approach are in good agreement with each other. The experimental data clearly demonstrated that acid/base chemistry in ILs is similar to that observed in molecular nonaqueous solvents; i.e., the relative strengths of the bases were in the right order and spaced (ΔpK(a)). It was also observed that the strength of N-bases is highly dependent on the anion of the ionic liquid; this observation indicates that pH-dependent reactions could be controlled by the appropriate choice of anion for bulk ILs or as an added co-ion to bulk IL.

Neotectonic and seismic activity in the Armorican and Cornubian Massifs: Regional stress field with glacio-isostatic influence?

Abstract In Brittany and SW England, evidence for low magnitude Quaternary seismicity can be found in sand pit exposures and beach sections. Deformation is especially well seen in alluvial and estuarine complexes resting on Late Pliocene sands or thick saprolite. The deformations are shallow, dominantly hydroplastic (high water-table) and led to overconsolidated sands, silts or clays. They reveal normal loading at micro (millimetric) and macro (decametric) scales as controlled by the local rheological properties of the sediments, by strikeslip faults associated with positive flower structures, folding, and intraformational water expulsion or hill slope sliding with reverse microfaulting. All the sites where these features occur, are in the vicinity of presumed active faults or steep slopes in highly fractured Proterozoic basement rocks indicating a possible shear zone. In most cases, these features are not associated with synsedimentary deformation, as strong superficial red soils are generally reworked by them. All these features are reworked by microfaulting after overconsolidation. Additional periglacial phenomena are superimposed on them and are often confused with them. Deformation occurred after the development of Holstenian peats (isotopic stage 11,400 ka BP) at Crozon (Brittany), after 317 ka BP (beginning of isotopic stage 9) in the Vilaine estuary, and in most other sites before the last rubified pedogenesis in the Middle Pleistocene (presumed isotopic stage 9). These dates correspond to the same episode that gave rise to the last main reactivation of the fossil cliff around 300 ka BP and to local uplift. One or several seismic clusters have taken place, probably due to delayed crustal rebound after a major glacial event (stage 10) and to rapid loading resulting from younger ice sheet growth (stage 8). Similar events occurred in late stage 7 and late stage 5. These events might have locally amplified the crustal deformation of the old Brittany and Cornubian Hercynian massifs resulting from a regional stress field especially from 400 ka to 200 ka BP. Stratigraphical, geodynamical and paleoclimatological implications are discussed.

Binding of nucleotides AMP and ATP to yeast phosphofructokinase:evidence for distinct catalytic and regulatory subunits.

Abstract The binding of ATP to yeast phosphofructokinase, as monitored by flow dialysis, is heterogeneous and is adequately described by assuming two independent classes of three binding sites each per enzyme molecule. Under similar conditions, the binding of 5′AMP is homogeneous and a binding stoichiometry of three 5′AMP molecules per enzyme molecule is evaluated. Displacement experiments show that only the ATP molecules bound to the first class of three tight binding sites are displaced by an excess of 5′AMP. Thus, these tight ATP binding sites can be identified as “regulatory sites” in agreement with kinetic data. Furthermore, molecular weight determinations and electrophoresis results are consistent with an heterologous α 3 β 3 structure of the enzyme oligomer. Therefore the present binding data suggest that yeast phosphofructokinase is constituted by three “catalytic” and three “regulatory” subunits.

Middle Pleistocene raised beach anomalies in the English Channel: regional and global stratigraphic implications

Abstract Palaeo-shore positions help to evidence long-term eustatic changes and assist in the understanding of tectonic movements at a regional scale. Raised beaches anomalies exist in the Channel region and may result from deformations induced by neotectonic or by glacio-isostasy. The aim of this paper is to re-analyse, within their geodynamic context in the Channel and Dover Strait regions, the stratigraphy and the datings of palaeo-shores of Middle and Upper Pleistocene ages. This sector of Europe is characterised by strong geological contrasts and is controlled by two main geological boundaries: in the north, the Variscan Overthrust (corresponding approximately to the position of the Dover Strait) and, in the south, the Northern Branch of the Southern Brittany shearing zone. These two boundaries border a domain which seems to behave rather homogeneously on a large scale under the control of plate tectonics. Today, shorelines are subsiding north and south of this ‘Channel’ region. Episodic uplift largely controlled the open or closed status of the Dover Strait, especially after the Messinian and Early Quaternary, by reactivating Variscan structures. After 400 ka, global cooling allowed supplementary deformations in the area to be induced by glacio-isostatic rebound and clustered seismic activity during the phase of ice sheet building. Evidence of eight different transgressions dated by ESR from Oxygen Isotopic Stage (OIS) 13 to the end of OIS 5 shows the complexity of the sea-level records in a region unstable for isostatic and neotectonic reasons. Due to glacio-isostatic depression, transgressions are possible in late glacial times as well as during full interglacials. Most platforms were initially cut, during the Late Miocene, and seem to have been re-trimmed several times, especially by shore ice rafting since OIS 9. Regionally, the sea apparently rose to about the same level in O.I. Stages 11, 9 and 7. Glacio-isostatic and glacio-eustatic relative displacements of the sea level together with background tectonic movements have modified coastal positions and have temporarily altered the intensity of tidal currents. The oldest shoreline deposits are preserved only in subsiding areas, controlled by the deep crustal pattern. Neotectonics related to Variscan structure reactivation still dominates glacio-isostatic deformation and basin subsidence. The OIS 7 positive anomaly seems related to a regional relaxation event.

Influence of phosphate on the allosteric behavior of yeast phosphofructokinase

Abstract Initial rate data obtained with purified yeast phosphofructokinase (PFK) show an ATP dependent kinetic cooperativity with respect to fructose-6-phosphate. In the presence of 25 mM phosphate, the cooperativity index (Hill number) is related to the half saturation concentration of fructose-6-phosphate as predicted by the concerted allosteric model in the case of a “K-system”. In the absence of phosphate, however, the kinetic behavior of yeast PFK is more complex and the cooperativity index is invariant with respect to the half saturation concentration of fructose-6-phosphate which is increased by ATP. In both cases, 5′AMP behaves as a strong activator of the enzyme. These kinetic data suggest that the two distinct functions of ATP as phosphate donnor and as allosteric inhibitor, respectively, are supported by different binding sites. These regulatory properties of yeast PFK are discussed in relation to glycolytic oscillations.

Octameric structure of yeast phosphofructokinase as determined by crosslinking with disuccinimidyl β‐hydromuconate

Yeast phosphofructo~nase (EC 2.7.1.1 l), a key enzyme in the regulation of the glycolytic pathway, is built up of an equal number of two types of subunits (Y and /3 which are immunologically distinct [l] and further differentiated by their relative susceptibility to proteolytic degradation in the presence of specific ligands [2]. Binding experiments [3,4] as well as functional properties of yeast phosphofructokinase [4,5] were indicative of a small number of interacting protomers (3 or 4) equal to only half the number of subunits constituting the enzyme oligomer. However, quaternary structure of yeast phosphofructokinase is still a matter of controversy since molecular weight determinations were unable to discriminate unambiguously between a hexameric and an octameric structure of the enzyme [4,6,7]. Thus, the major evidence for an octameric structure was obtained from small-angle X-ray scattering [S] and from crosshnking experiments with dimethyl suberimidate [9]. However, these data could not rule out an eventual hexameric structure for yeast phosphofructokinase. Indeed, the interpretation of X-ray diffusion data relies heavily on the assumption of an arbitrary chosen shape of the subunits. Moreover, no polymeric forms higher than tetramer

Studies on the structure of yeast phosphofructokinase

In this paper, we describe an efficient procedure for the purification of yeast phosphofructokinase. This procedure eliminates any time delay and enables to obtain an enzyme with minimum proteolytic alterations. The molecular weights of the oligomeric enzyme and of its constitutive subunits were both evaluated by means of several independent methods. However, the accuracy of each measurement was not sufficient to discriminate between an hexameric and an octameric structure of the enzyme oligomer. On the other hand, crosslinking experiments demonstrated the octameric structure of yeast phosphofructokinase. Obviously, some methods of molecular weight determination have led to erroneous results. In particular, our experiments show that the reliability of molecular weight determinations performed by gel filtration of native proteins must be considered with caution.

A tectonic inversion by steps during the Cenozoic : the Dover Strait.

The Boulonnais is a dome incised by a former marine gulf inset into a zone of tectonic inversion from the Middle Eocene, which was already partly excavated at least at the Upper Eocene. New sedimentological and paleopedological data obtained within the Boulonnais, completed with old seismic profiles, allow a better understanding of the inversion process which developed step by step. The initial breaching probably took place in the late Eocene. The Dover Strait was probably opened during the Lutetian, a part of the Oligocene and the late Neogene. Oligocene and Pliocene faunal assemblages are identical on both sides of the Strait. It was closed again for tectonic and eustatic reasons in the early Quaternary and reopen subsequently just before the Last Interglacial. The opening is related to the evolution of the Western Channel and of its paleovalley system. The inversion of the Variscan front accommodates most of the shortening induced by the Pyrenean Orogen on the western border of the European plate. The inversion of the Dover Strait region is almost synchronous with those of other basins of the Channel and North Sea areas. Tectonic, geomorphologic and climatic implications of this dynamic are discussed within the western European context.