Extraction and immunochemical characterization of delta sleep-inducing peptide-like material from the porcine pituitary and adrenal gland.
The naturally occurring forms of delta sleep-inducing peptide (DSIP) are not fully identified. In the present study, porcine pituitaries and adrenal glands were extracted in water, saline or acid under various conditions and immunoreactive DSIP (IR-DSIP) quantified by radioimmunoassay. The highest concentrations were measured in anterior pituitary extracts (40.8 +/- 2.6 ng/g tissue weight) recovered using water with aprotinin. However, high performance liquid chromatography (HPLC) indicated degradation of hydrophobic forms of IR-DSIP in water extracts. Extraction in acetic acid including C18 Sep-Pak purification resulted in an elution profile of IR-DSIP in adrenal extracts with a major peak coeluting with synthetic DSIP [DSIP(1-9)], whereas anterior pituitary extract showed material of higher hydrophobicity. Approximately 30% of IR-DSIP in anterior pituitary as well as in adrenal gland extracts seemed to be glucosylated, as based on concanavalin A chromatography. One of the DSIP-immunoreactive components by immunoblotting (molecular mass 25 kDa) was identified in both pituitary and adrenal gland extracts. In conclusion, several chromatographically distinct forms of IR-DSIP are present in the porcine pituitary and adrenal gland. IR material eluting as DSIP(1-9) is present in adrenal gland extract. The procedure and solution used for tissue extraction seem to be essential in order to obtain reliable elution positions on HPLC.
Department of Psychiatry, University of Lund, Sweden.
A
A
Bjartell
F
F
Sundler
R
R
Ekman
D000311:Adrenal Glands / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D002845:Chromatography; D003701:Delta Sleep-Inducing Peptide / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D015151:Immunoblotting; D007120:Immunochemistry; D008969:Molecular Sequence Data; D010902:Pituitary Gland / Q000737:chemistry*; D011863:Radioimmunoassay; D013552:Swine
D003701:Delta Sleep-Inducing Peptide