Spontaneous Pulsation of Peptide Microstructures in an Abiotic Liquid System.

We report observations of pulsating peptide formation and depeptidization in 70% aqueous acetonitrile solutions of l-Pro-l-Phe and l-Cys, resulting in the oscillatory appearance and disappearance of solid masses of microfibers and microspheres, respectively. We monitor the concentration changes of the monomeric amino acids by high-performance liquid chromatography. The concentration of all amino acid solutions used is 1.0 mg mL(-1), due to solubility limitations in 70% aqueous acetonitrile. The nonlinear concentration changes of l-Pro, l-Phe and l-Cys, and the amounts of the main peptidization products observed within our monitoring periods (for l-Pro-l-Phe, 250 h, and for l-Cys, 70 h) are typically from several to 20% of the original monomer concentrations. We follow the formation and decay of the insoluble peptides by turbidimetry. We also investigate the materials formed using scanning electron microscopy and mass spectrometry. We carry out numerical simulations on a simple model that reflects the main features of spontaneous pulsation of peptide fiber or sphere formation in this abiotic liquid system.

Dynamics of Spontaneous Peptidization of l-, d- and dl-Serine in an Abiotic Solution as Investigated with Use of TLC-Densitometry and the Auxiliary Chromatographic Techniques.

From our earlier investigations, it comes out that proteinogenic amino acids can undergo spontaneous oscillatory reactions of chiral inversion and peptidization. l-Serine (l-Ser) is an important proteinogenic amino acid with many vital functions in human and mammalian organisms, e.g., it is responsible for good condition of the nervous cell membranes. It undergoes spontaneous oscillatory processes of chiral inversion and peptidization, and the goal of this study was to compare the dynamics of its peptidization with that of d-Ser and dl-Ser (racemate). The main analytical technique used in our experiment was TLC-densitometry, and the auxiliary chromatographic techniques were HPLC-evaporative light scattering detector and LC-MS. The results obtained witness to the differences in peptidization dynamics of the two Ser enantiomers (l and d) and of the racemic mixture thereof (dl). It was shown that dl-Ser characterizes with the higher, and l- and d-Ser with the lower peptidization yields.

On Spontaneously Pulsating Proline-Phenylalanine Peptide Microfibers.

Earlier, we have collected an experimental evidence showing that low molecular weight chiral carboxylic acids (amino acids included) can spontaneously undergo an oscillatory chiral conversion and an oscillatory condensation in abiotic aqueous and non-aqueous liquid systems, stored for certain amount of time under mild external conditions. These earlier findings are summarized in the introductory part of this study. In the second part, a preliminary report is given on spontaneous pulsation of peptide microfibers in the aged proline-phenylalanine (Pro-Phe) solution in 70% aqueous acetonitrile. The experimental evidence originates from a number of advanced analytical techniques. In view of our earlier and present findings, a presumption is made that the mechanism of spontaneous pulsation (formation and decay) of Pro-Phe microfibers is directly related to the oscillatory chiral conversion and oscillatory peptidization. The entity of the discussed results pointing out to spontaneous and uncontrolled instability of peptide structures might be a bad prognostic for employing such structures in nanobiotechnology.

HPLC monitoring of spontaneous non-linear peptidization dynamics of selected amino acids in solution.

This is our new study in a series of publications devoted to exploration of applicability of high-performance liquid chromatography (HPLC) to providing answers to difficult questions from the area of the reaction kinetics and mechanisms with non-linear reactions. Although an excellent analytical performance of HPLC is an indisputable fact, so far its performance as a tool in the kinetic and mechanistic studies has been tested to a lesser extent. In our earlier studies, spontaneous peptidization dynamics of amino acids in solution was demonstrated by means of HPLC upon a few amino acid examples, and on that basis a theoretical model has been developed, anticipating an interdependence of dynamics on chemical structures of amino acids involved. In order to expand the spectrum of experimentally investigated amino acid cases, in this study we present the results valid for three novel amino acids of significant life sciences importance, which differ in terms of peptidization dynamics. Experimental evidence originates from the achiral HPLC with the evaporative light scattering detection and MS detection. A conclusion is drawn that different spontaneous peptidization dynamics of amino acids may significantly influence chemical composition of proteins encountered in living organisms. Hence, a need emerges for systematic physicochemical studies on spontaneous non-linear peptidization dynamics of proteinogenic amino acids in liquid abiotic (but also in the biotic) systems.

Condensation dynamics of the L-Pro-L-Phe and L-Hyp-L-Phe binary mixtures in solution.

We employ the achiral liquid chromatography with diode array, evaporative light scattering and mass spectrometric detection (HPLC-DAD, HPLC-ELSD and LC-MS) to assess structural instability (understood as spontaneous oscillatory chiral conversion and spontaneous oscillatory condensation) of the two pairs of amino acids, L-proline-L-phenylalanine (L-Pro-L-Phe) and L-hydroxyproline-L-phenylalanine (L-Hyp-L-Phe), in aqueous acetonitrile. In our earlier studies, we managed to demonstrate that single amino acids in aqueous and non-aqueous solutions undergo spontaneous oscillatory chiral conversion and oscillatory condensation. We also investigated condensation in the binary L-Pro-L-Hyp mixture in aqueous solution, and proposed a theoretical model to explain the specific dynamics of this process, which involves mutual catalytic effects of the two amino acids. In this study, we demonstrate oscillatory instability with the other two amino acid pairs in the organic-aqueous solution and reflect on the dynamics of condensation in the investigated cases. The choice of L-Pro and L-Hyp is due to their important role as building blocks of collagen, which is omnipresent in the connective tissues of mammals, and largely responsible for tissue architecture and strength. L-Phe is one of the 20 exogenous amino acids and is a building block of the majority of naturally occurring proteins.