Separation of Acetylated Neomycins B and C by Paper Chromatography

Pan, S.C.; Dutcher, James D.

doi:10.1021/ac60113a017

Cited by 166 publications

(39 citation statements)

References 10 publications

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“…These could be detected by ultraviolet absorption and by the chlorination/starch/potassium iodide method! 19 1, but the ninhydrin reaction was very faint. At pH 2.1 'comets' were observed with mobilities to the cathode of 1.75 (|3) and 1.60 (7) that of e-dinitrophenyllysine.…”

Section: Resultsmentioning

confidence: 96%

The Isolation of the Urogastrones — Inhibitors of Gastric Acid Secretion — from Human Urine

Gregory¹,

Willshire²

1975

Hoppe-Seyler´s Zeitschrift für physiologische Chemie

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It has been known for over thirty years that extracts of human urine could cause inhibition of gastric acid secretion. The active principle was called urogastrone and this has now been isolated using a series of twelve simple stages of partition, gel and ion-exchange chromatography. In fact two products were obtained, each in a yield of less than l mg per 1000 / urine, which represented an overall recovery of 3-5%. These materials were biologically indistinguishable, causing inhibition of gastric acid secretion in a variety of circumstances at doses of less than 1 Mg/kg. The purified urogastrones were found to be acidic polypeptides composed of 53 and 52 amino acid residues with three internal disulphide bonds, and they differed by only one arginine residue. Neither threonine nor phenylalanine residues were present in the urogastrones. Isolierung von Urogastronen (Hemmstoffen für die Magensäuresekretion) aus menschlichem UrinZusammenfassung: Seit über dreißig Jahren ist bekannt, daß Extrakte aus menschlichem Urin die Magensäuresekretion hemmen können. Das Urogastron genannte aktive Prinzip wurde in 12 einfachen Reinigungsstufen durch Verteilungs-, Gelund lonenaustausch-Chromatographie isoliert. Es wurden zwei Produkte in einer Ausbeute von jeweils weniger als l mg/1000 / Urin erhalten (Gesamt-Wiederfindung 3 -5%). Diese Substanzen lassen sich biologisch nicht voneinander unterscheiden (getestet in ihrer die Magensäuresekre-tion hemmenden Wirkung bei Dosen von weniger als l Mg/kg unter verschiedenen Bedingungen).

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Section: Resultsmentioning

confidence: 96%

The Isolation of the Urogastrones — Inhibitors of Gastric Acid Secretion — from Human Urine

Gregory¹,

Willshire²

1975

Hoppe-Seyler´s Zeitschrift für physiologische Chemie

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“…The hydroxyproline peptide present in this portion of the column did not react with Ninhydrin, isatin, or dinitrofluorobenzene (DNFB), suggesting absence of a free terminal amino or imino acid. When a portion of the complete fraction was subjected to paper electrophoresis in one dimension, followed by chromatography in the second dimension (solvent A) (fingerprinting), the hypro peptide could be located in an area which gave a positive reaction for peptide when sprayed with the Rydon and Smith reagents, as modified by Pan and Dutcher (17). The peptide behaved as a neutral component under the electrophoretic conditions described above, and migrated with an Rf similar to leucine in solvent A.…”

Section: Resultsmentioning

confidence: 99%

Urinary Hydroxyproline Peptides*

Meilman¹,

Urivetzky²,

Rapoport³

1963

J. Clin. Invest.

110

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“…Solvent systems were the following: (a) phenol: water, 5: 1; (b) n-butanol: glacial acetic acid: water, 18: 2: 5 and 4:1: 5 (15); and (c) isobutanol: 3.3%o ammonium hydroxide, 5: 2. Chromatograms were stained for dipeptide with 0.2% isatin in n-butanol and for the diketopiperazine of prolylhydroxyproline with the starch-iodide spray of Pan and Dutcher (16).…”

Section: Methods Preparation Of 'Ic-labeled Gelatin Two Male Weanlingmentioning

confidence: 99%

The quantitative relationship of urinary peptide hydroxyproline excretion to collagen degradation

Weiss¹,

Klein²

1969

J. Clin. Invest.

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A B S T R A C r To determine the quantitative relationship of urinary hydroxyproline peptide excretion to collagen breakdown, known quantities of radioactive hydroxyproline peptides were administered to unlabeled animals and excertion of radioactivity in respiratory carbon dioxide, urine, and feces was measured. The major routes of excretion of collagen peptide metabolites were respiratory carbon dioxide (75%) and urine, as hydroxyproline-containing peptides (25%).Since the predominant urine hydroxyproline peptide linkage is prolyl-hydroxyproline, L-prOlyl-L-hydroxyproline-3H was administered to unlabeled animals. Greater than 80% of the administered dipeptide was excreted in urine, suggesting that this peptide linkage is not hydrolyzed to a significant extent in vivo.These data suggest that urinary hydroxyproline excretion is a "fairly" sensitive indicator of collagen breakdown and can be used at the clinical level to quantitate changes in collagen breakdown. creted in urine, about 97% in the form of peptides and 3% as the free imino acid (3). The dietary intake of gelatin has resulted in a marked increase of urinary excretion of bound hydroxyproline (3,4). The amino acid composition of urinary hydroxyproline peptides before (5, 6) and after (6) gelatin feeding in normal subjects suggests that urinary hydroxyproline peptides are endproducts of collagen degradation. The predominant urinary hydroxyproline peptides, prolyl-hydroxyproline and glycyl-prolyl-hydroxyproline, contain amino acid sequences known to occur frequently in the collagen molecule (7). To further implicate collagen breakdown as the source of urinary hydroxyproline peptides are the observations that excretion increases or decreases in a number of clinical conditions where an increase or decrease in collagen breakdown would be expected (8)(9)(10)(11). INTRODUCTIONUrinary hydroxyproline peptide excretion would be most useful in the analysis of diseases of connective tissues at the clinical level if it represents a quantitatively significant and fairly constant fraction of the hydroxyproline released by collagen degradation. Since urinary hydroxyproline excretion is almost entirely in the form of peptides, its quantitative importance would appear to depend on what proportion of the peptides released by the degradation of collagen is excreted in urine, what proportion is further degraded to carbon dioxide, and whether this is a fairly constant relationship.The present investigation approaches the question of the quantitative significance of urinary hydroxyproline peptide excretion by techniques similar to those used in the study of plasma proteins (12, 13). Known quantities of radioactive hydroxyproline peptides were administered to unlabeled animals and the distribution of excretion in urine, respiratory carbon dioxide, and feces was measured. This approach permits an estimation of the quantitative significance of each of these pathways from the distribution of excretion of radioactivity. The Journal of Clinical Investigation Volume 48 1969 1These s...

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Separation of Acetylated Neomycins B and C by Paper Chromatography

Cited by 166 publications

References 10 publications

The Isolation of the Urogastrones — Inhibitors of Gastric Acid Secretion — from Human Urine

The Isolation of the Urogastrones — Inhibitors of Gastric Acid Secretion — from Human Urine

Urinary Hydroxyproline Peptides*

The quantitative relationship of urinary peptide hydroxyproline excretion to collagen degradation

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