Cloning and expression of a cDNA coding for a human monocyte-derived plasminogen activator inhibitor
Autor: | T. M. Antalis, P. L. Devine, N. H. Goss, M. A. Clark, Ross W. Stephens, G. Schevzov, Thomas M. Barnes, P. Tolstoshev, P. R. Lehrbach |
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Rok vydání: | 1988 |
Předmět: |
Ovalbumin
Recombinant Fusion Proteins Molecular Sequence Data Monocytes Cell Line Rapid amplification of cDNA ends Complementary DNA Sequence Homology Nucleic Acid Coding region Animals Humans Protease Inhibitors Amino Acid Sequence Peptide sequence Glycoproteins Multidisciplinary biology Base Sequence cDNA library Nucleic acid sequence DNA Molecular biology Plasminogen Inactivators Biochemistry Multigene Family biology.protein Plasminogen activator Chickens Research Article |
Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 85(4) |
ISSN: | 0027-8424 |
Popis: | 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. |
Databáze: | OpenAIRE |
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