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Elizabeth L Ostler

Publications and source records attributed to Elizabeth L Ostler.

8 recordsLinked to original sources
PubMed2007

Evidence that the mechanism of antibody-catalysed hydrolysis of arylcarbamates can be determined by the structure of the immunogen used to elicit the catalytic antibody.

A kinetically homogeneous anti-phosphate catalytic antibody preparation was shown to catalyse the hydrolysis of a series of O-aryl N-methyl carbamates containing various substituents in the 4-position of the O-phenyl group. The specific nature of the antibody catalysis was demonstrated by the adherence of these reactions to the Michaelis-Menten equation, the complete inhibition by a hapten analogue, and the failure of the antibody to catalyse the hydrolysis of the 2-nitrophenyl analogue of the 4-nitrophenylcarbamate substrate. Hammett sigma-rho analysis suggests that both the non-catalysed and antibody-catalysed reactions proceed by mechanisms in which development of the aryloxyanion of the leaving group is well advanced in the transition state of the rate-determining step. This is probably the ElcB (elimination-addition) mechanism for the non-catalysed reaction, but for the antibody-catalysed reaction might be either ElcB or B(Ac)2 (addition-elimination), in which the elimination of the aryloxy group from the tetrahedral intermediate has become rate-determining. This result provides evidence of the dominance of recognition of phenolate ion character in the phosphate hapten in the elicitation process, and is discussed in connection with data from the literature that suggest a B(Ac)2 mechanism, with rate-determining formation of the tetrahedral intermediate for the hydrolysis of carbamate substrates catalysed by an antibody elicited by a phosphonamidate hapten in which phenolate anion character is minimized. The present paper contributes to the growing awareness that small differences in the structure of haptens can produce large differences in catalytic characteristics.

Animals↗
PubMed2004

What can progeroid syndromes tell us about human aging?

Human genetic diseases that resemble accelerated aging provide useful models for gerontologists. They combine known single-gene mutations with deficits in selected tissues that are reminiscent of changes seen during normal aging. Here, we describe recent progress toward linking molecular and cellular changes with the phenotype seen in two of these disorders. One in particular, Werner syndrome, provides evidence to support the hypothesis that the senescence of somatic cells may be a causal agent of normal aging.

Aging↗
PubMed2004

Evidence for 'lock and key' character in an anti-phosphonate hydrolytic antibody catalytic site augmented by non-reaction centre recognition: variation in substrate selectivity between an anti-phosphonate antibody, an anti-phosphate antibody and two hydrolytic enzymes.

The substrate selectivities of an anti-phosphonate and an anti-phosphate kinetically homogeneous polyclonal catalytic antibody preparation and two hydrolytic enzymes were compared by using hapten-analogous and truncated carbonate and ester substrates each containing a 4-nitrophenolate leaving group. Syntheses of the truncated substrates devoid of recognition features in the non-leaving group parts of the substrates are reported. The relatively high kinetic selectivity of the more active anti-phosphonate antibody preparation is considered to depend on a relatively rigid catalytic site with substantial reaction centre specificity together with other important recognition interactions with the extended non-leaving group part of the substrate. In contrast, the less catalytically active, more flexible anti-phosphate antibody exhibits much lower kinetic selectivity for the substrate reaction centre comparable with that of the hydrolytic enzymes with activity much less dependent on recognition interactions with the non-leaving group part of the substrate. The ways in which haptenic flexibility and IgG architecture might contribute to the differential kinetic selectivities are indicated.

Animals↗
PubMed2003

Improvement in hydrolytic antibody activity by change in haptenic structure from phosphate to phosphonate with retention of a common leaving-group determinant: evidence for the 'flexibility' hypothesis.

To investigate the hypothesis that decreased hapten flexibility may lead to increased catalytic antibody activity, we used two closely related immunogens differing only in the flexibility of the atomic framework around the structural motif of the haptens, analogous to the reaction centre of the corresponding substrates. Identical leaving-group determinants in the haptens and identical leaving groups in the substrates removed the ambiguity inherent in some data reported in the literature. Anti-phosphate and anti-phosphonate kinetically homogeneous polyclonal catalytic antibody preparations were compared by using carbonate and ester substrates respectively, each containing a 4-nitrophenolate leaving group. Synthetic routes to a new phosphonate hapten and new ester substrate were developed. The kinetic advantage of the more rigid anti-phosphonate/ester system was demonstrated at pH 8.0 by a 13-fold advantage in k(cat)/k(non-cat) and a 100-fold advantage in the proficiency constant, k(cat)/k (non-cat) x K(m). Despite these differences, the pH-dependences of the kinetic and binding characteristics and the results of chemical modification studies suggest closely similar catalytic mechanisms. The possible origin of the kinetic advantage of the more rigid hapten/substrate system is discussed.

Antibodies, Catalytic↗
PubMed2002

A rat monoclonal antibody that catalyses the hydrolysis of a nitrophenyl-beta-lactam.

We report the first example of a monoclonal antibody-catalysed hydrolysis of a beta-lactam where the antibodies were generated by a simple transition-state analogue. A rat monoclonal antibody (1/91c/4d/26) generated by using an acyclic 4-nitrophenylphosphate immunogen catalysed the hydrolysis of corresponding 4-nitrophenyl carbonates but, more importantly, also catalysed the hydrolysis of N-(4-nitrophenyl)-azetidinone at pH 8 with k(cat)=8.7 x 10(-6)s(-1) and K(M)=35 microM. This is the first example of a rat monoclonal catalytic antibody.

Animals↗
PubMed2002

Polyclonal catalytic antibodies.

Polyclonal catalytic antibodies offer advantages in the evaluation of immunogens and in the ease of production of large quantities of antibodies. They comprise the entire immune response of an animal to an immunogen where monoclonals represent a subset. Polyclonal antibodies are consequently particularly suitable for evaluating catalytic antibody responses generated by different haptens or a group of structurally related haptens. The authors reported the first polyclonal catalytic antibodies in 1990. An unexpected finding is that polyclonal catalytic antibodies show single-site kinetic behaviour, i.e. whatever structural heterogeneity exists, the kinetic behaviour is homogeneous. Many groups worldwide have since published work in this area. Three groups are prominent. The authors' group, a group based in Austin, Texas, and led by Iverson, and a Shanghai group. The authors' group works with sheep antibodies and has published mechanistic studies and, more recently, specificity studies that revealed the catalysis of a beta-lactam. Most of this work over a 10-year period was performed by using a single bleed from a single sheep, which gives an indication of the ease of production and utility of such catalytic antibodies. Iverson's group works with rabbit antibodies and has published much work on the evaluation of catalytic antibodies generated by structurally related haptens. The Shanghai group has concentrated on the studies of polyclonal catalytic antibodies for electrocyclic reactions where any danger of contaminating enzymes is reduced. The use of immunisation to generate therapeutically useful catalytic antibodies (necessarily polyclonal) in a host animal is an attractive target. Several groups are working towards this. The authors themselves have published in this area and promising recent studies come from a group working in France on immunisation to protect against nerve agents and a Texas group (based in Houston) that has demonstrated active immunisation to generate antibodies that catalyse the hydrolysis of a carbamate insecticide.

Animals↗
PubMedDate not supplied

Can we say that senescent cells cause ageing?

Replicative senescence, the irreversible loss of proliferative capacity, is a common feature of somatic cells derived from many different species. The molecular mechanisms controlling senescence in mammals, and especially in humans, have now been substantively elucidated. However, to date, attempts to link the senescence of cells with the ageing of the organisms they comprise has not met with any similar degree of success, largely due to a lack of systematic investigation and the absence of the necessary biochemical tools. This review will summarise current data linking replicative senescence and organismal ageing. It will also suggest some essential tests of the cell senescence hypothesis and some necessary ground work which must be carried out before such tests can be fruitfully performed. It will not discuss the detailed molecular 'clockwork' controlling the decision to exit the cell cycle irreversibly because this is covered by other authors in this special issue.

Aging↗