Isolation and characterization of a corticotropin-releasing hormone-like peptide from human placenta.
Immunoreactive CRH was detected in extracts of human term placentae [5.2 +/- 0.8 (+/- SE) pmol/g wet wt; n = 9]. Molecular sieve chromatography revealed three size classes of immunoreactive CRH. The major species eluted with the Kav of synthetic rat CRH; the minor species had apparent mol wt (MW) of 18,000 and 8,000. A placental CRH-(1-41)-sized peptide was isolated by fractional acetone precipitation, molecular sieve chromatography, and sequential reverse phase high performance liquid chromatography steps. This peptide had the same chromatographic behavior as did rat CRH in all high performance liquid chromatographic isolation steps as well as the same UV absorbance to immunoreactive CRH ratio after the final purification step. Purified placental CRH stimulated ACTH release from anterior pituitary tissue in a dose-dependent manner and was equipotent with synthetic rat CRH. Partial sequencing indicated that 32 amino acids of this peptide are identical to those of rat and human CRH (sequence deduced from genomic sequence), and comparative peptide mapping with rat CRH provided further evidence that the placental CRH-like peptide is very homologous if not identical to CRH. The high mol wt placental CRH fractions also were partially purified by acetone precipitation, immune affinity chromatography, and gel filtration. Neither of these materials [big form (MW, 18,000) or intermediate form (MWr, 8,000)] stimulated ACTH release from rat pituitary tissue in vitro. Limited trypsin digestion of the highest MW CRH, followed by gel filtration analysis, resulted in conversion to the smaller [8,000 MW-sized and CRH-(1-41)-sized] forms. The detection of a CRH-like peptide in placenta together with our previous demonstration of plasma immunoreactive CRH in pregnant women suggest that the placenta synthesizes and secretes CRH into the maternal circulation.
Sasaki
A
A
Department of Internal Medicine, Wakayanagi Hospital, Miyagi, Japan.
Tempst
P
P
Liotta
A S
AS
Margioris
A N
AN
Hood
L E
LE
Kent
S B
SB
Sato
S
S
Shinkawa
O
O
Yoshinaga
K
K
Krieger
D T
DT
D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003346:Corticotropin-Releasing Hormone / Q000302:isolation & purification* / Q000502:physiology; D005260:Female; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000302:isolation & purification*; D010920:Placenta / Q000032:analysis*; D011247:Pregnancy; D011863:Radioimmunoassay; D014357:Trypsin
D010455:Peptides; D003346:Corticotropin-Releasing Hormone; D014357:Trypsin