Structural Characterization of O‐ and C‐Glycosylating Variants of the Landomycin Glycosyltransferase LanGT2

The structures of the O‐glycosyltransferase LanGT2 and the engineered, C? C bond‐forming variant LanGT2S8Ac show how the replacement of a single loop can change the functionality of the enzyme. Crystal structures of the enzymes in complex with a nonhydrolyzable nucleotide‐sugar analogue revealed that there is a conformational transition to create the binding sites for the aglycon substrate. This induced‐fit transition was explored by molecular docking experiments with various aglycon substrates.     

Using molecular dynamics and quantum mechanics calculations to model fluorescence observables

We provide a critical examination of two different methods for generating a donor-acceptor electronic coupling trajectory from a molecular dynamics (MD) trajectory and three methods for sampling that coupling trajectory, allowing the modeling of experimental observables directly from the MD simulation. In the first coupling method we perform a single quantum-mechanical (QM) calculation to characterize the excited state behavior, specifically the transition dipole moment, of the fluorescent probe, which is then mapped onto the configuration space sampled by MD. We then utilize these transition dipoles within the ideal dipole approximation (IDA) to determine the electronic coupling between the probes that mediates the transfer of energy. In the second method we perform a QM calculation on each snapshot and use the complete transition densities to calculate the electronic coupling without need for the IDA. The resulting coupling trajectories are then sampled using three methods ranging from an independent sampling of each trajectory point (the independent snapshot method) to a Markov chain treatment that accounts for the dynamics of the coupling in determining effective rates. The results show that the IDA significantly overestimates the energy transfer rate (by a factor of 2.6) during the portions of the trajectory in which the probes are close to each other. Comparison of the sampling methods shows that the Markov chain approach yields more realistic observables at both high and low FRET efficiencies. Differences between the three sampling methods are discussed in terms of the different mechanisms for averaging over structural dynamics in the system. Convergence of the Markov chain method is carefully examined. Together, the methods for estimating coupling and for sampling the coupling provide a mechanism for directly connecting the structural dynamics modeled by MD with fluorescence observables determined through FRET experiments.     

Atmosph?rische Luft

Ohne Zusammenfassung     

A frequency stabilized,cw dye laser for high resolution spectroscopy

A single frequency, jet stream cw dye laser spectrometer locked to a stable high finesse optical reference cavity is described. This system is capable of maintaining a frequency stability of ± 1 MHz/min and a continuous scan of up to 6 GHz.     

The internal sphere condition and the capillary problem

Sunto In un importante lavoro [1], M. Emmer ha dimostrato l’esistenza di superfici di equilibrio capillari, definite in domini con frontiera Lipschitziana, quando la costante di Lipschitz L e l’angolo di contatto γ soddisfano la relazione L<|tg γ|. Questa condizione, che è noto essere necessaria in generale, può essere troppo restrittiva in alcuni casi particolari. In questo lavoro la condizione di Emmer è sostituita da una ? condizione di sfera interna ?, che conduce ad una larga classe di domini che non rientrano nel risultato di Emmer. L’esiestenza di una soluzione è dimostrata anche nel caso γ=0, che non può essere trattato con il metodo di Emmer.

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Entrata in Redazione il 26 agosto 1975.

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This work was initiated while the former author was at Universit?t Bonn and at Università di Genova, and the latter author at Université de Paris VI as a Fellow of the Deutsche Forschungsgemeinschaft.