Abstract
A sleep-promoting substance, Factor S, has been extracted and purified from large volumes of human urine. Urinary Factor S is a small glycopeptide; amino acid-amino sugar analyses of the purified material revealed a substance composed of glutamic acid, alanine, diaminopimelic acid, and muramic acid in molar ratios of 2:2:1:1. The active glycopeptide resembles bacterial peptidoglycans but the composition suggests that it is not simply of adventitious origin. Other reasons for this conclusion are also given. Infusions into the lateral ventricle of the brain of about 5 pmol/kg of body weight induce a 50% increase in slow wave sleep in rabbits. The excess sleep is normal as judged by electrophysiological and behavioral criteria; it resembles the deep sleep that occurs when animals are allowed to sleep following prolonged sleep deprivation.
Highlights
Amino acids released by acid hydrolysis of several purified samples of urinary Factor S
Amino acids and amino sugars released by mild acid hydrolysis of reported that intraventricular infusion of very small amounts purified urinary Factor S
This seems unlikely because: (a)previously we showed that urinary Factor S passes through an Amicon
Summary
Materials logical and behavioral criteria; it resembles the deep Human urine was obtained from healthy male adults. The lyophilized product from step B was dissolved in an appropriate volume of 50 mM NH1-acetate buffer, pH 5.0 (1ml buffer/liter of original urine) This solution was applied to a DEAE-Sephadex column (bed volume of 66 ml/100 liters of original urine). The lyophilized product from step D was taken up in this buffer (0.4 ml/100 liters of original urine) and applied to the SP-Sephadex column. The column was developed by washing with Beckman microcolumn lithium citrate buffer, pH 2.83(0.24 N LiOH, 0.16 N citric acid, 0.25% thiodiglycol, 16 ml of concentrated HCl/liter [12]), and separatefractions each representing about 1/12 of a bed volume were collected. In the case of the SP-Sephadex step, the fractions on each side of those fractionsthat contained sleep-promoting activity were examined each time thesecolumns were developed, to serve as blanks These were all negative in the biological assay. Instrument parameters were optimized for either single label counting ([‘4C]sucrose, step Bo)r for double label counting ([3H]cysteic acid and [“C]aspartic acid, step E)
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