Purification, characterization, and cDNA cloning of a Kunitz-type proteinase inhibitor secreted by the porcine uterus.
The porcine uterus synthesizes a proteinase inhibitor (M(r) 14,000) under the influence of progesterone that is relatively specific for plasmin and trypsin, but that also has weak affinity for chymotrypsin. Several isoforms of this uterine plasmin/trypsin inhibitor were purified by a procedure whose final two steps involved affinity chromatography on immobilized chymotrypsin and cation exchange chromatography. Amino-terminal sequencing showed that at least three of the isoforms were closely related. An oligonucleotide probe based on the protein sequence was used to identify a cDNA that contained an open reading frame coding for a mature protein (M(r) 10,295) of 93 amino acids. The inhibitor had a well defined, but unique, Kunitz domain of 64 residues at its amino terminus that shared 67% sequence identity to bovine pancreatic trypsin inhibitor. Its P1 residue was arginine rather than lysine. Northern analysis showed the presence of a single mRNA species (700 bases) that in adult female pigs appeared to be confined to the uterus. During pregnancy, UPTI mRNA expression was high until Day 30 and decreased significantly thereafter. By contrast, uteroferrin mRNA reached maximal concentrations in late pregnancy. These data are consistent with an earlier hypothesis that the inhibitor serves to neutralize the activities of one or more serine proteinases generated by the proliferating trophoblast during the formation of the noninvasive placenta of the pig.
Stallings-Mann
M L
ML
Department of Animal Sciences, University of Missouri, Columbia 65211.
Burke
M G
MG
Trout
W E
WE
Roberts
R M
RM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D010455:Peptides; D010940:Plant Proteins; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013552:Swine; D014361:Trypsin Inhibitors / Q000235:genetics / Q000378:metabolism*; D014599:Uterus / Q000201:enzymology* / Q000378:metabolism
D018076:DNA, Complementary; C053865:Kunitz-type protease inhibitor, plant; D010455:Peptides; D010940:Plant Proteins; D012333:RNA, Messenger; D014361:Trypsin Inhibitors