NMR analysis of synthetic human serum albumin alpha-helix 28 identifies structural distortion upon amadori modification

The non-enzymatic reaction between reducing sugars and long-lived proteins in vivo results in the formation of glycation and advanced glycation end products, which alter the properties of proteins including charge, helicity, and their tendency to aggregate. Such protein modifications are linked with various pathologies associated with the general aging process such as Alzheimer disease and the long-term complications of diabetes. Although it has been suggested that glycation and advanced glycation end products altered protein structure and helicity, little structural data and information currently exist on whether or not glycation does indeed influence or change local protein secondary structure. We have addressed this problem using a model helical peptide system containing a di-lysine motif derived from human serum albumin. We have shown that, in the presence of 50 mM glucose and at 37 degrees C, one of the lysine residues in the di-lysine motif within this peptide is preferentially glycated. Using NMR analysis, we have confirmed that the synthetic peptide constituting this helix does indeed form a alpha-helix in solution in the presence of 30% trifluoroethanol. Glycation of the model peptide resulted in the distortion of the alpha-helix, forcing the region of the helix around the site of glycation to adopt a 3(10) helical structure. This is the first reported evidence that glycation can influence or change local protein secondary structure. The implications and biological significance of such structural changes on protein function are discussed

\u3csup\u3e13\u3c/sup\u3eC NMR Investigation of Nonenzymatic Glucosylation of Protein

Nonenzymatic glucosylation of protein is initiated by the reversible condensation of glucose in its open chain form with the amino groups on the protein. The initial product is an aldimine (Schiff base) which cyclizes to the glycosylamine derivative. The aldimine can undergo a slow Amadori rearrangement to yield the relatively stable ketoamine adduct which is structurally analogous to fructose. 13C NMR has been used to characterize these early products of nonenzymatic glucosylation, using RNase A as a model protein. C-1 of the beta-pyranose anomer of the glycosylamine was identified at 88.8 ppm in the spectrum of RNase glucosylated approximately 1:1 with D-[1-13C]glucose. C-1 of the Amadori product was also apparent in this spectrum, resonating as a pair of intense peaks at 52.7 and 53.1 ppm. The anomeric (C-2) resonances of the Amadori adduct were seen in the spectrum of RNase glucosylated approximately 1:1 with [U-13C]glucose. This spectrum was interpreted by comparison to the spectra of reference compounds: D-fructose, fructose-glycine, N alpha-formyl-N epsilon-fructose-lysine, and glucosylated poly-L-lysine. In the protein spectrum, the most intense of the C-2 resonances was that of the beta-fructopyranose anomer at 95.8 ppm. The alpha- and beta-fructofuranose anomers were also observed at 101.7 and 99.2 ppm, respectively. One unidentified signal in the anomeric region was observed in the spectra of poly-L-lysine and RNase, both glucosylated with [U-13C]glucose; no comparable resonances were observed in the spectra of the model compounds

Spray-dried olive mill wastewater reduces Maillard reaction in cookies model system

The network of the Maillard reaction can be influenced by the presence of polyphenols. In this paper, we evaluated the ability of secoiridoids to interact with asparagine and lysine tuning the formation of dietary advanced glycation end-products (d-AGEs), dicarbonyls and acrylamide. Olive oil mill wastewater polyphenol powders (OMWP) were added to glucose and lysine or asparagine in silica model systems to mimic water activity present in cookies. Results revealed that acrylamide, Amadori compounds and N-ε-carboxyethyllysine (CEL) were reduced to 50%, after 13 min at 180°C; for the reduction of N-ε-carboxymethyllysine (CML), secoiridoids were effective only in model systems with the addition of acacia fiber and maltodextrin as coating agents. In cookies, OMWP at three different concentrations decreased the concentration of protein bound Amadori compounds, CML, CEL and dicarbonyls. Acrylamide and 5-hydroxymethylfurfural were reduced to 60% and 76% respectively, highlighting the ability of secoiridoids-based functional ingredients in controlling d-AGEs formation

Evolution of protein bound Maillard reaction end-products and free Amadori compounds in low lactose milk in presence of fructosamine oxidase I

Thermal treatments and storage influence milk quality, particularly in low lactose milk as the higher concentration of reducing sugars can lead to the increased formation of the Maillard reaction products (MRPs). The control of the Amadori products (APs) formation is the key step to mitigate the Maillard reaction (MR) in milk. The use of fructosamine oxidases, (Faox) provided promising results. In this paper, the effects of Faox I were evaluated by monitoring the concentration of free and bound MRPs in low lactose milk during shelf life. Results showed that the enzyme reduced the formation of protein-bound MRPs down to 79% after six days at 37 °C. Faox I lowered the glycation of almost all the free amino acids resulting effective on basic and polar amino acids. Data here reported corroborate previous findings on the potentiality of Faox enzymes in controlling the early stage of the MR in foods

Asymptotic profile and Morse index of nodal radial solutions to the H\'enon problem

We compute the Morse index of nodal radial solutions to the H\'enon problem $$\left\{\begin{array}{ll} -\Delta u = |x|^{\alpha}|u|^{p-1} u \qquad & \text{ in } B, \newline u= 0 & \text{ on } \partial B, \end{array} \right.$$ where $B$ stands for the unit ball in ${\mathbb R}^N$ in dimension $N\ge 3$, $\alpha>0$ and $p$ is near at the threshold exponent for existence of solutions $p_{\alpha}=\frac{N+2+2\alpha}{N-2}$, obtaining that \begin{align*} m(u_p) & = m \sum\limits_{j=0}^{1+\left[{\alpha}/{2}\right]} N_j \quad & \mbox{ if $\alpha$ is not an even integer, or} \newline m(u_p)& = m\sum\limits_{j=0}^{ \alpha /2} N_j + (m-1) N_{1+\alpha/ 2} & \mbox{ if $\alpha$ is an even number.} \end{align*} Here $N_j$ denotes the multiplicity of the spherical harmonics of order $j$. The computation builds on a characterization of the Morse index by means of a one dimensional singular eigenvalue problem, and is carried out by a detailed picture of the asymptotic behavior of both the solution and the singular eigenvalues and eigenfunctions. In particular it is shown that nodal radial solutions have multiple blow-up at the origin, where each node converges (up to a suitable rescaling) to the bubble shaped solution of a limit problem. As side outcome we see that solutions are nondegenerate for $p$ near at $p_{\alpha}$, and we give an existence result in perturbed balls.Comment: 47 page

1-Dibenzylamino-1-de­oxy-4,5-O-isopropyl­idene-β-d-fructopyran­ose

The title compound C23H29NO5, synthesized by the Amadori rearrangement of α-d-glucose with dibenzyl­amine and the ketalization, is shown to be a β-anomer. The fructopyran­ose ring adopts a chair conformation. The two benzene rings form a dihedral angle of 68.9 (1)°. In the crystal, non–classical inter­molecular C—H⋯O hydrogen bonds link the mol­ecules into a three–dimensional network

Nonradial sign changing solutions to Lane Emden equation

In this paper we prove the existence of continua of nonradial solutions for the Lane-Emden equation. In a first result we show that there are infinitely many global continua detaching from the curve of radial solutions with any prescribed number of nodal zones. Next, using the fixed point index in cone, we produce nonradial solutions with a new type of symmetry. This result also applies to solutions with fixed signed, showing that the set of solutions to the Lane Emden problem has a very rich and complex structure.Comment: 13 p

Pharmacologic approaches against advanced glycation end products (ages) in diabetic cardiovascular disease

Advanced Glycation End-Products (AGEs) are signaling proteins associated to several vascular and neurological complications in diabetic and non-diabetic patients. AGEs proved to be a marker of negative outcome in both diabetes management and surgical procedures in these patients. The reported role of AGEs prompted the development of pharmacological inhibitors of their effects, giving rise to a number of both preclinical and clinical studies. Clinical trials with anti-AGEs drugs have been gradually developed and this review aimed to summarize most relevant reports

Bifurcation and symmetry breaking for the H\'enon equation

In this paper we consider the H\'enon problem in a ball. We prove the existence of (at least) one branch of nonradial solutions that bifurcate from the radial ones and that this branch is unbounded