Cloning and expression of a cDNA coding for a human monocyte-derived plasminogen activator inhibitor.
Human monocyte-derived plasminogen activator inhibitor (mPAI-2) was purified to homogeneity from the U937 cell line and partially sequenced. Oligonucleotide probes derived from this sequence were used to screen a cDNA library prepared from U937 cells. One positive clone was sequenced and contained most of the coding sequence as well as a long incomplete 3' untranslated region (1112 base pairs). This cDNA sequence was shown to encode mPAI-2 by hybrid-select translation. A cDNA clone encoding the remainder of the mPAI-2 mRNA was obtained by primer extension of U937 poly(A)+ RNA using a probe complementary to the mPAI-2 coding region. The coding sequence for mPAI-2 was placed under the control of the lambda PL promoter, and the protein expressed in Escherichia coli formed a complex with urokinase that could be detected immunologically. By nucleotide sequence analysis, mPAI-2 cDNA encodes a protein containing 415 amino acids with a predicted unglycosylated Mr of 46,543. The predicted amino acid sequence of mPAI-2 is very similar to placental PAI-2 (3 amino acid differences) and shows extensive homology with members of the serine protease inhibitor (serpin) superfamily. mPAI-2 was found to be more homologous to ovalbumin (37%) than the endothelial plasminogen activator inhibitor, PAI-1 (26%). Like ovalbumin, mPAI-2 appears to have no typical amino-terminal signal sequence. The 3' untranslated region of the mPAI-2 cDNA contains a putative regulatory sequence that has been associated with the inflammatory mediators.
Antalis
T M
TM
Biotechnology Australia Pty. Ltd., Roseville, New South Wales, Australia.
Clark
M A
MA
Barnes
T
T
Lehrbach
P R
PR
Devine
P L
PL
Schevzov
G
G
Goss
N H
NH
Stephens
R W
RW
Tolstoshev
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002645:Chickens; D004247:DNA / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D009000:Monocytes / Q000032:analysis; D005810:Multigene Family; D010047:Ovalbumin / Q000235:genetics; D015849:Plasminogen Inactivators; D011480:Protease Inhibitors / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
D006023:Glycoproteins; D015849:Plasminogen Inactivators; D011480:Protease Inhibitors; D011993:Recombinant Fusion Proteins; D010047:Ovalbumin; D004247:DNA