Different posttranslational processing of human preprothyrotropin-releasing hormone in the human placenta and hypothalamus.
We scrutinized the possible existence of human prepro-THR messenger RNA (mRNA) and of its posttranslational processing products in the human placenta. Human placental mRNA of preproTRH was found to have a single species identical to that predicted from the hypothalamic mRNA, and could be reverse transcribed to complementary DNA (cDNA) encoding preproTRH in a size similar to the hypothalamic counterpart by means of reverse-transcriptase-polymerase chain reaction. Five different intervening peptides, designated human TRH-associated peptide (hTAP) [hTAP-1, preproTRH(90-111); hTAP-2, preproTRH(120-132); hTAP-3, preproTRH(141-149); hTAP-4, preproTRH(158-183); hTAP-5, preproTRH(192-224)], and a TRH precursor comprising the TRH progenitor sequence (octa-TRH) were synthesized, and six different antisera raised against individual peptides were used to develop specific RIA systems. Significant concentrations of hTAP-5 and octa-TRH-like immunoreactivities were quantitated in acid extracts of human placentae. In human hypothalamic extracts, immunoreactivities of hTAP-3, hTAP-4, hTAP-5, and octa-TRH, were apparently detected. Chromotographic analysis showed a single peak corresponding to each authentic peptide in RIA systems of hTAPs detected. In placentae, a single peak of octa-TRH-like substance was observed, and two octa-TRH-like substances with different molecular weights detected in hypothalami. The present data indicate that unique posttranslational processing steps of human preproTRH differ in human placentae and hypothalami, and that the human tissues involve profound amounts of several preproTRH-related peptides which do not comprise the TRH progenitor sequence.
Mori
M
M
First Department of Medicine, Gunma University, School of Medicine, Maebashi, Japan.
Yamada
M
M
Satoh
T
T
Murakami
M
M
Iriuchijima
T
T
Kobayashi
I
I
D000368:Aged; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D004247:DNA / Q000378:metabolism; D005260:Female; D006801:Humans; D007031:Hypothalamus / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D010920:Placenta / Q000378:metabolism*; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000378:metabolism; D011863:Radioimmunoassay; D013973:Thyrotropin-Releasing Hormone / Q000235:genetics / Q000378:metabolism*
D015335:Molecular Probes; D010446:Peptide Fragments; D011498:Protein Precursors; D012333:RNA, Messenger; C063596:prepro-thyrotropin-releasing hormone; D013973:Thyrotropin-Releasing Hormone; D004247:DNA