Carlos Duque-Daza

Tandem mass spectrometry for the study of glyoxal-derived advanced glycation end-products (AGEs) in peptides.

RATIONALE The post-translational modification known as glycation affects the physiological properties of peptides and proteins. Glycation is particularly important during hyperglycaemia where α-dicarbonyl compounds are generated. These compounds react with proteins to generate α-dicarbonyl-derived glycation products, which are correlated with diabetic complications such as nephropathy, retinopathy, and neuropathy, among others. One of these α-dicarbonyl compounds is ethanedial, also known as glyoxal. Thereby, glyoxal binding to protein/peptides is studied by electron capture dissociation (ECD) and collisionally activated dissociation (CAD). METHODS Acetylated and non-acetylated undecapeptides containing one lysine and one arginine susceptible of glycation were reacted with glyoxal under pseudo-physiological and MeOH/H2O (50:50) conditions. Two types of glyoxal-derived AGEs were fragmented by ECD and CAD using 12 Tesla Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS). RESULTS Reaction with glyoxal under different reaction conditions showed the addition of C2O and C2H2O2, which corresponded to a net increase on the peptide mass of 39.9949 Da and 58.0055 Da, respectively. The binding site was assigned within an error <1 ppm, using ECD and CAD. The results indicated that both types of glyoxal-derived AGEs are formed at the side chain of arginine located in position 3. CONCLUSIONS Types and binding sites of glyoxal-derived AGEs were investigated in peptides containing one arginine-one lysine using FTICRMS. Two net mass additions to the mass of the peptide were assigned as C2O and C2H2O2, which were located at the arginine side chain. In addition, these mass additions (C2O and C2H2O2) observed in the peptides were unaffected by different reaction conditions.

Unexpected Crosslinking and Diglycation as Advanced Glycation End-Products from Glyoxal

AbstractGlyoxal-derived advanced glycation end-products (AGEs) are formed in physiological systems affecting protein/peptide function and structure. These AGEs are generated during aging and chronic diseases such as diabetes and are considered arginine glycating agents. Thus, the study of glyoxal-derived AGEs in lysine residues and amino acid competition is addressed here using acetylated and non-acetylated undecapeptides, with one arginine and one lysine residue available for glycation. Tandem mass spectrometry results from a Fourier transform ion cyclotron resonance mass spectrometer showed glycated species at both the arginine and lysine residues. One species with the mass addition of 116.01096 Da is formed at the arginine residue. A possible structure is proposed to explain this finding (Nδ-[2-(dihydroxymethyl)-2H,3aH,4H,6aH-[1, 3]dioxolo[5,6-d]imidazolin-5-yl]-L-ornithine-derived AGE). The second species corresponded to intramolecular crosslink involving the lysine residue and its presence is checked with ion-mobility mass spectrometry. Graphical Abstractᅟ

Efficient Time-Step Coupling For Hybrid Continuum/Molecular Modelling of Unsteady Micro-Scale Gas Flows

In this paper we describe a numerical method for the efficient time-accurate coupling of hybrid continuum/molecular micro gas flow solvers. Hybrid approaches are commonly used when non-equilibrium effects in the flow field are spatially localized; in these regions a more accurate, but typically more expensive, solution procedure is adopted. Although this can greatly increase efficiency in steady flows, in unsteady flows the evolution of the solution as a whole is restricted by the maximum time step allowed by the molecular-based/kinetic model; numerically speaking, this is a stiff problem. In the method presented in this paper we exploit time-scale separation, when it exists, to partially decouple the temporal evolution of the two parts of the hybrid model. This affords major computational savings. The method is a modified/extended version of the seamless heterogeneous multiscale method (SHMM). Our approach allows multiple micro steps (molecular steps) before coupling with the macro (continuum) solver: we call this a multi-step SHMM. This maintains the main advantages of SHMM (computational speed-up and flexible application) while improving on accuracy and greatly reducing the number of continuum computations and instances of coupling required. The improved accuracy of the multi-step SHMM is demonstrated for two canonical one-dimensional transient flows (oscillatory Poiseuille and oscillatory Couette flow) and for rarefied-gas oscillatory Poiseuille flow.

Numerical solution of the Falkner-Skan equation using third-order and high-order-compact finite difference schemes

We present a computational study of the solution of the Falkner-Skan equation (a third-order boundary value problem arising in boundary-layer theory) using high-order and high-order-compact finite differences schemes. There are a number of previously reported solution approaches that adopt a reduced-order system of equations, and numerical methods such as: shooting, Taylor series, Runge-Kutta and other semi-analytic methods. Interestingly, though, methods that solve the original non-reduced third-order equation directly are absent from the literature. Two high-order schemes are presented using both explicit (third-order) and implicit compact- difference (fourth-order) formulations on a semi-infinite domain; to our knowledge this is the first time that high-order finite difference schemes are presented to find numerical solutions to the non-reduced-order Falkner- Skan equation directly. This approach maintains the simplicity of Taylor-series coefficient matching methods, avoiding complicated numerical algorithms, and in turn presents valuable information about the numerical behaviour of the equation. The accuracy and effectiveness of this approach is established by comparison with published data for accelerating, constant and decelerating flows; excellent agreement is observed. In general, the numerical behaviour of formulations that seek an optimum physical domain size (for a given computational grid) is discussed. Based on new insight into such methods, an alternative optimisation procedure is proposed that should increase the range of initial seed points for which convergence can be achieved. Keywords: laminar boundary layer, similarity analysis, high-order-compact finite differences