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.
Bjartell
A
A
Department of Psychiatry, University of Lund, Sweden.
Sundler
F
F
Ekman
R
R
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