Purification, measurement, and tissue distribution of a dansyl-derivatized glycopeptide from low-molecular weight follicle-stimulating hormone-inhibitor-containing fractions of porcine follicular fluid.
We have used dansyl chloride (5-dimethylamino-1-naphthalenesulfonyl choloride) to form dansyl derivatives of amine-containing compounds in follicular fluid or highly purified fractions containing a low molecular weight (MW) inhibitor of follicle-stimulating hormone (FSH) binding to receptor (FSH-BI). This approach allowed sensitive detection of the derivatives based on their fluorescent properties. By taking advantage of the hydrophobic nature of the dansyl group, a dansyl derivative (RF = 0.15) identified in low MW FSH-BI preparations was purified from porcine follicular fluid. Based on chromatographic criteria using four different systems (thin-layer chromatography [TLC] and high performance liquid chromatography), the derivatized factor (D15) that was purified appeared to be homogeneous. A direct, chemical assay was developed for quantification of D15 from follicular fluid or tissue extracts. The highest concentration (153 ng/mg) of D15 was found in ovarian tissue of adult rats, lesser amounts were observed in kidney and liver tissues (93 and 62 ng/mg, respectively) and even less in diaphram and heart tissues (5 and 0.5 ng/mg, respectively). High concentrations of D15 were observed in derivatized extracts of tests from immature rats in which approximately twice as much D15 was found in Leydig cells (241 ng/mg) as in seminiferous tubules (136 ng/mg). In porcine ovarian tissue, granulosa cells from large follicles and corpora lutea (69 and 91 ng/mg, respectively) contained at least 4-fold higher concentrations than follicle wall tissue (14 ng/ml). Relative concentrations of D15 material were also determined in pools of bovine follicular fluid previously shown to contain low MW FSH-BI.(ABSTRACT TRUNCATED AT 250 WORDS)
Sluss
P M
PM
Department of Biochemistry, Albany Medical College, New York 12208.
Branca
A A
AA
Ford
J J
JJ
Krishnan
K A
KA
Reichert
L E
LE
D000818:Animals; D001826:Body Fluids / Q000032:analysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003619:Dansyl Compounds; D005260:Female; D005453:Fluorescence; D005640:Follicle Stimulating Hormone / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D006020:Glycopeptides / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D008297:Male; D008970:Molecular Weight; D006080:Ovarian Follicle / Q000032:analysis*; D011247:Pregnancy; D051381:Rats; D011962:Receptors, FSH / Q000378:metabolism; D013552:Swine
D003619:Dansyl Compounds; D006020:Glycopeptides; D011962:Receptors, FSH; D005640:Follicle Stimulating Hormone; C029038:dansyl chloride