Proline from hydroxyproline

The chromatograms stained with ninhydrin are immersed in the reagent solution for 1 s or sprayed evenly with it and then placed in the free half of a twin-trough chamber containing 25% ammonia solution. Apart from proline and hydroxyproline, which yield yellow copper complexes, all the amino acids yield reddish-colored chromatogram zones [3],... 

The tendencies of the amino acids to stabilize or destabilize a-helices are different in typical proteins than in polyamino acids. The occurrence of the common amino acids in helices is summarized in Table 6.1. Notably, proline (and hydroxyproline) act as helix breakers due to their unique structure, which fixes the value of the —N—C bond angle. Helices can be formed from either... 

Animal glue is a complex colloidal mixture of proteins. The related gelatins are also complex heterogeneous mixtures of proteins. They are strongly hydrophilic and rich in the amino acids glycine, proline, lysine, hydroxyproline and hydroxylysine. Casein is a phosphoprotein obtained from the milk of mammals. 

Other workers began to study the structure of gramicidin. Christensen and coworkers12 isolated crystalline tryptophane and leucine from a hydrolysate. They found no evidence for a fatty acid component and established that phenylalanine, proline and hydroxyproline were absent from a hydrolysate. These workers isolated alanine diox-pyridate from a hydrolysate and also established that gramicidin contained a compound with vicinal hydroxy and amino groups. They speculated that this compound might be serine or isoserine and proposed that gramicidin contains two tryptophane, 2 leucine, 2 or 3 alanine and 1 hydroxyamino residues or a multiple of this composition. 

Plaquet et al. (PI) found in the urine of rachitic children peptides consisting of proline, hydroxyproline, and glycine, which they believed to be the products of collagen degradation. Two similar peptides containing considerable amounts of proline and hydroxyproline were isolated from the urine of a patient with rheumatoid arthritis by Mechanic et al. (Ml). One of these peptides consisted of three proline, two hydroxyproline, and nine glutamic acid residues, the second one consisted of four proline, four hydroxyproline, and one glutamic acid residues. The N-terminal amino acid in the first peptide was demonstrated to be hydroxyproline. 

Ninhydrin (triketohydrindene hydrate) reacts with an amino acid when heated under acidic conditions (pH 3-4) to produce ammonia, carbon dioxide and a blue-purple complex. This reaction forms the basis of many widely used methods (Figure 10.11). One mole of carbon dioxide is liberated from each mole of amino acid, exceptions being the dicarboxylic amino acids, which produce two moles of carbon dioxide, and the a-imino acids, proline and hydroxyproline, which do not produce carbon dioxide. Although this formed the basis of a gasometric technique, colorimetric methods are now the most common. 

Sowden et al. [4] also did detailed amino acid and amino sugar analyses of the soils from the different dimatic regions. The following amino acids were determined acidic amino acids aspartic and glutamic acids basic amino acids arginine, histidine, lysine and ornithine neutral amino adds, phenylalanine, tyrosine, glycine, alanine, valine, leudne, isoleudne, serine, threonine, proline and hydroxyproline ... 

Proline and hydroxyproline differ from the other amino acids in that they contain a secondary amino group in a pyrrolidine ring therefore, they do not produce aminoketones and Strecker aldehydes in the reaction with dicarbonyls, ffowever, nitrogen heterocyclics are produced, including 1-pyrroline, pyrrolidine, 2-acetyl-1-pyrroline and 2-acetyltetrahydropyridine (Scheme 12.5) [19]. 

Gl (MW >5000). Most of the pyrolysis fragments found in this fraction are derived from the three general classes just mentioned. Pyrrole and methylpyrrole originate from proteinaceous material such as polypeptides as well as from single amino acids such as proline and hydroxyproline. A quantitative relationship between amino acid hydrolyzable content and pyrrole abundance was established by Bracewell (20) for some Scottish brown forest soils, and such a correlation probably could be established for water. 

The non-alkaloid derived organocatalysts 13a-e - readily accessible from proline and hydroxyproline, respectively - were reported by Uozomi et al. (Scheme 13.7) [17]. Of the five compounds, 13b and 13e performed best. In the presence of 100 mol% 13e, the methanolytic desymmetrization of cyclic meso anhydrides was found to proceed with up to 89% ee. 

In 1899 Fischer turned his attention to the study of proteins, wishing to understand their chemical structures. It was known at that time that proteins were composed of amino acids, and thirteen naturally occurring ones were identified. Fischer was able to isolate via the hydrolysis of proteins three additional naturally occurring amino acids valine, proline, and hydroxyproline. Amino acids exhibit stereoisomerism, and Fischer was able to separate individual forms from mixtures of stereoisomers for several of these compounds. 

Scheme 5.3 Formation of proline- and hydroxyproline-specific heterocyclic volatiles from [1 -13C glucose via 3-deoxyglucosone223...

Table I. Mole Ratio of Proline to Hydroxyproline in Collagen Isolated from Granuloma of Normal and Scorbutic Guinea Pigs0...

Table III. Incorporation of Proline into Collagen Proline and Hydroxyproline of Granulomas from Normal and Scorbutic Guinea Pigs...

It is conceivable that the reduced formation of tritiated water observed in scurvy could be due to a general reduction in protein synthesis, but this would not explain the marked differences between the specific activities of proline and hydroxyproline observed, nor would it explain observations in our laboratory that glycine and tyrosine are incorporated at rates similar to that of incorporation of proline into the collagen of granuloma obtained from scorbutic animals. 

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