Although it is generally accepted that amino acids were present on the prebiotic Earth, the mechanism by which α-amino acids were condensed into polypeptides before the emergence of enzymes remains unsolved. Here, we demonstrate a prebiotically plausible mechanism for peptide (amide) bond formation that is enabled by α-hydroxy acids, which were likely present along with amino acids on the early Earth. Together, α-hydroxy acids and α-amino acids form depsipeptides—oligomers with a combination of ester and amide linkages—in model prebiotic reactions that are driven by wet–cool/dry–hot cycles. Through a combination of ester–amide bond exchange and ester bond hydrolysis, depsipeptides are enriched with amino acids over time. These results support a long-standing hypothesis that peptides might have arisen from ester-based precursors.

Unlike traditional drift-tube ion mobility-mass spectrometry, traveling-wave ion mobility-mass spectrometry typically requires calibration in order to generate collision cross section (CCS) values. Although this has received a significant amount of attention for positive-ion mode analysis, little attention has been paid for CCS calibration in negative ion mode. Here, we provide drift-tube CCS values for [M - H](-) ions of two calibrant series, polyalanine and polymalic acid, and evaluate both types of calibrants in terms of the accuracy and precision of the traveling-wave ion mobility CCS values that they produce.

Wet–dry cycling is widely regarded as a means of driving condensation reactions under prebiotic conditions to generate mixtures of prospective biopolymers. A criticism of this model is its reliance on unpredictable rehydration events, like rainstorms. Here, we report the ability of deliquescent minerals to mediate the oligomerization of glycine during iterative wet–dry cycles. The reaction mixtures evaporate to dryness at high temperatures and spontaneously reacquire water vapor to form aqueous solutions at low temperatures. Deliquescent mixtures can foster yields of oligomerization over ten-fold higher than non-deliquescent controls. The deliquescent mixtures tightly regulate their moisture content, which is crucial, as too little water precludes dissolution of the reactants while too much water favors hydrolysis over condensation. The model also suggests a potential reason why life evolved to favor the enrichment of potassium: so living systems could acquire and retain sufficient water to serve as a solvent for biochemical reactions.

The rise of peptides with secondary structures and functions would have been a key step in the chemical evolution which led to life. As with modern biology, amino acid sequence would have been a primary determinant of peptide structure and activity in an originsof-life scenario. It is a commonly held hypothesis that unique functional sequences would have emerged from a diverse soup of proto-peptides, yet there is a lack of experimental data in support of this. Whereas the majority of studies in the field focus on peptides containing only one or two types of amino acids, here we used modern mass spectrometry (MS)-based techniques to separate and sequence de novo proto-peptides containing broader combinations of prebiotically plausible monomers. Using a dry-wet environmental cycling protocol, hundreds of proto-peptide sequences were formed over a mere 4 d of reaction. Sequence homology diagrams were constructed to compare experimental and theoretical sequence spaces of tetrameric proto-peptides. MS-based analyses such as this will be increasingly necessary as origins-of-life researchers move toward systems-level investigations of prebiotic chemistry. . Soon after, Fox and Harada began to explore the formation of peptides from amino acids via condensation at high temperatures (2). These reactions were facilitated by proportionally higher concentrations of amino acids with acidic side chains (e.g., aspartic acid, D) and resulted in the production of "proteinoids," condensation products containing covalent cross-links not found in coded proteins. In their 1960 manuscript, Fox and Harada speculated about the diversity of proteinoid sequences, yet conceded that "a complete answer to the question of whether the amino acid residues are distributed in a random or other arrangement may require a complete assignment of residues in one molecular species," a task beyond the analytical capabilities of the time (3).In subsequent years, the proteinoid concept was displaced by the hypothesis that either RNA or proto-RNA gave rise to life (4-6). Nevertheless, to this day, condensation reactions of amino acids are thought to have played a key role in a potentially symbiotic proto-nucleic acid and proto-peptide world (7,8). Peptide condensation studies at temperatures lower than those of Fox and Harada, or with the aid of chemical agents, confirmed that abiotic production of peptides was indeed possible, but chain lengths were generally limited to dimers and trimers with low yields (9). In recent studies, this length barrier has been surpassed, and certain proto-peptides have been shown to form aggregate structures (10, 11). Nevertheless, the majority of proto-peptide studies have been limited in scope, typically containing only one or two types of amino acid monomer.We recently introduced a model prebiotic pathway for peptide formation based on ester-amide exchange reactions between α-amino acids and α-hydroxy acids (12), amino acid structural analogs found in meteorites and model prebiotic reactions (13, 14) (Fig. 1A). Subjecting ...

Traditional methods for deriving computationally-generated collision cross sections for comparisons with ion mobility-mass spectrometry data require 3-dimensional energy-minimized structures and are often time consuming, preventing high throughput implementation. Here, we introduce a method to predict ion mobility collision cross sections of lipids and peptide analogs important in prebiotic chemistry and other fields. Using less than 100 2-D molecular descriptors this approach resulted in prediction errors of less than 2%.

Although it is generally accepted that amino acids were present on the prebiotic Earth, the mechanism by which a-amino acids were condensed into polypeptides before the emergence of enzymes remains unsolved. Here,w ed emonstrate aprebiotically plausible mechanismfor peptide (amide) bond formation that is enabled by a-hydroxy acids,which were likely present along with amino acids on the early Earth. Together, a-hydroxy acids and a-amino acids form depsipeptides-oligomers with ac ombination of ester and amide linkages-in model prebiotic reactions that are driven by wet-cool/dry-hot cycles.T hrough ac ombination of esteramide bond exchange and ester bond hydrolysis,depsipeptides are enriched with amino acids over time.These results support al ong-standing hypothesis that peptides might have arisen from ester-based precursors.

Profiling and imaging of cholesterol and its precursors by mass spectrometry (MS) are important in a number of cholesterol biosynthesis disorders, such as in Smith-Lemli-Opitz syndrome (SLOS), where 7-dehydrocholesterol (7-DHC) is accumulated in affected individuals. SLOS is caused by defects in the enzyme that reduces 7-DHC to cholesterol. However, analysis of sterols is challenging because these hydrophobic olefins are difficult to ionize for MS detection. We report here sputtered silver matrix-assisted laser desorption/ionization (MALDI)-ion mobility-MS (IM-MS) analysis of cholesterol and 7-DHC. In comparison with liquid-based AgNO3 and colloidal Ag nanoparticle (AgNP), sputtered silver NP (10–25 nm) provided the lowest limits-of-detection based on the silver coordinated [cholesterol+Ag]+ and [7-DHC+Ag]+ signals while minimizing dehydrogenation products ([M+Ag-2H]+). When analyzing human fibroblasts that were directly grown on poly-L-lysine-coated ITO glass plates with this technique, in situ, the 7-DHC/cholesterol ratios for both control and SLOS human fibroblasts are readily obtained. The m/z of 491 (specific for [7-DHC+107Ag]+) and 495 (specific for [cholesterol+109Ag]+) were subsequently imaged using MALDI-IM-MS. MS images were co-registered with optical images of the cells for metabolic ratio determination. From these comparisons, ratios of 7-DHC/cholesterol for SLOS human fibroblasts are distinctly higher than in control human fibroblasts. Thus, this strategy demonstrates the utility for diagnosing/assaying the severity of cholesterol biosynthesis disorders in vitro.

Purified methylenedianiline (MDA) regioisomers were structurally characterized and differentiated using tandem mass spectrometry (MS/MS), ion mobility-mass spectrometry (IM-MS), and IM-MS/MS in conjunction with computational methods. It was determined that protonation sites on the isomers can vary depending on the position of amino groups, and the resulting protonation sites play a role in the gas-phase stability of the isomer. We also observed differences in the relative distributions of protonated conformations depending on experimental conditions and instrumentation, which is consistent with previous studies on aniline in the gas phase. This work demonstrates the utility of a multifaceted approach for the study of isobaric species and elucidates why previous MDA studies may have been unable to detect and/or differentiate certain isomers. Such analysis may prove useful in the characterization of larger MDA multimeric species, industrial MDA mixtures, and methylene diphenyl diisocyanate (MDI) mixtures used in polyurethane synthesis.