Identification of a serpin-enzyme complex receptor on human hepatoma cells and human monocytes

David H. Perlmutter, George I. Glover, Meheryar Rivetna, Charles S. Schasteen, Robert Fallon

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

Formation of the covalently stabilized complex of α1-antitrypsin (α1-AT) with neutrophil elastase, the archetype of serine proteinase inhibitor serpin-enzyme complexes, is associated with structural rearrangement of the α1-AT molecule and hydrolysis of a reactive-site peptide bond. An ≈ 4-kDa carboxyl-terminal cleavage fragment is generated. α1-AT-elastase complexes are biologically active, possessing chemotactic activity and mediating increases in expression of the α1-AT gene in human monocytes and macrophages. This suggested that structural rearrangement of the α1-AT molecule, during formation of a complex with elastase, exposes a domain that is recognized by a specific cell surface receptor or receptors. To test this hypothesis, the known three-dimensional structure of α1,-AT and comparisons of the primary structures of the serpins were used to select a potentially exteriorly exposed and highly conserved region in the complexed form of α1-AT as a candidate ligand (carboxyl-terminal fragment, amino acids 359-374). We show here that synthetic peptides based on the sequence of this region bind specifically and anturably to human hepatoma cells and human monocytes (Kd = 4.0 × 10-8 M, 4.5 × 105 plasma membrane receptors per cell) and mediate increases in synthesis of α1-AT. Binding of peptide 105Y (Ser-Ile-Pro-Pro-Glu-Val-Lys-Phe-Asn-Lys-Pro-Phe-Val-Tyr-Leu-Ile) is blocked by α1-AT-elastase complexes, antithrombin III (AT Ill)-thrombin complexes, α1-antichymotrypsin (α1-ACT)-cathepsin G complexes, and, to a ser extent, complement component C1 inhibitor-C1s complexes, but not by the corresponding native proteins. Binding of peptide 105Y is also blocked by peptides with sequence corresponding to carboxyl-terminal fragments of the serpins AT III and α1-ACT, but not by peptides having the sequence of the extreme amino terminus of α1-AT. The results also show that peptide 105Y inhibits binding of 125I-labeled α1-AT-aducstase complexes. Thus, these studies demonstrate an abundant, relatively high-affinity cell surface receptor which recognizes serpin-enzyme complexes (SEC receptor). This receptor is capable of modulating the production of at least one of the serpins, α1-AT. Since the ligand specificity is similar to that previously described for in vivo clearance of serpin-enzyme complexes, the SEC receptor may also be involved in the clearance of certain serpin-enzyme complexes.

Original languageEnglish (US)
Pages (from-to)3753-3757
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number10
StatePublished - 1990
Externally publishedYes

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Serpins
Monocytes
Hepatocellular Carcinoma
Peptides
Pancreatic Elastase
Cell Surface Receptors
Cathepsin G
Complement C1
Ligands
Leukocyte Elastase
Serine Proteinase Inhibitors
Antithrombin III
Enzymes
serpin-enzyme complex receptor
Thrombin
Catalytic Domain
Hydrolysis
Macrophages
Cell Membrane
Amino Acids

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Identification of a serpin-enzyme complex receptor on human hepatoma cells and human monocytes. / Perlmutter, David H.; Glover, George I.; Rivetna, Meheryar; Schasteen, Charles S.; Fallon, Robert.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 10, 1990, p. 3753-3757.

Research output: Contribution to journalArticle

Perlmutter, David H. ; Glover, George I. ; Rivetna, Meheryar ; Schasteen, Charles S. ; Fallon, Robert. / Identification of a serpin-enzyme complex receptor on human hepatoma cells and human monocytes. In: Proceedings of the National Academy of Sciences of the United States of America. 1990 ; Vol. 87, No. 10. pp. 3753-3757.
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abstract = "Formation of the covalently stabilized complex of α1-antitrypsin (α1-AT) with neutrophil elastase, the archetype of serine proteinase inhibitor serpin-enzyme complexes, is associated with structural rearrangement of the α1-AT molecule and hydrolysis of a reactive-site peptide bond. An ≈ 4-kDa carboxyl-terminal cleavage fragment is generated. α1-AT-elastase complexes are biologically active, possessing chemotactic activity and mediating increases in expression of the α1-AT gene in human monocytes and macrophages. This suggested that structural rearrangement of the α1-AT molecule, during formation of a complex with elastase, exposes a domain that is recognized by a specific cell surface receptor or receptors. To test this hypothesis, the known three-dimensional structure of α1,-AT and comparisons of the primary structures of the serpins were used to select a potentially exteriorly exposed and highly conserved region in the complexed form of α1-AT as a candidate ligand (carboxyl-terminal fragment, amino acids 359-374). We show here that synthetic peptides based on the sequence of this region bind specifically and anturably to human hepatoma cells and human monocytes (Kd = 4.0 × 10-8 M, 4.5 × 105 plasma membrane receptors per cell) and mediate increases in synthesis of α1-AT. Binding of peptide 105Y (Ser-Ile-Pro-Pro-Glu-Val-Lys-Phe-Asn-Lys-Pro-Phe-Val-Tyr-Leu-Ile) is blocked by α1-AT-elastase complexes, antithrombin III (AT Ill)-thrombin complexes, α1-antichymotrypsin (α1-ACT)-cathepsin G complexes, and, to a ser extent, complement component C1 inhibitor-C1s complexes, but not by the corresponding native proteins. Binding of peptide 105Y is also blocked by peptides with sequence corresponding to carboxyl-terminal fragments of the serpins AT III and α1-ACT, but not by peptides having the sequence of the extreme amino terminus of α1-AT. The results also show that peptide 105Y inhibits binding of 125I-labeled α1-AT-aducstase complexes. Thus, these studies demonstrate an abundant, relatively high-affinity cell surface receptor which recognizes serpin-enzyme complexes (SEC receptor). This receptor is capable of modulating the production of at least one of the serpins, α1-AT. Since the ligand specificity is similar to that previously described for in vivo clearance of serpin-enzyme complexes, the SEC receptor may also be involved in the clearance of certain serpin-enzyme complexes.",
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T1 - Identification of a serpin-enzyme complex receptor on human hepatoma cells and human monocytes

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AU - Glover, George I.

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AU - Schasteen, Charles S.

AU - Fallon, Robert

PY - 1990

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N2 - Formation of the covalently stabilized complex of α1-antitrypsin (α1-AT) with neutrophil elastase, the archetype of serine proteinase inhibitor serpin-enzyme complexes, is associated with structural rearrangement of the α1-AT molecule and hydrolysis of a reactive-site peptide bond. An ≈ 4-kDa carboxyl-terminal cleavage fragment is generated. α1-AT-elastase complexes are biologically active, possessing chemotactic activity and mediating increases in expression of the α1-AT gene in human monocytes and macrophages. This suggested that structural rearrangement of the α1-AT molecule, during formation of a complex with elastase, exposes a domain that is recognized by a specific cell surface receptor or receptors. To test this hypothesis, the known three-dimensional structure of α1,-AT and comparisons of the primary structures of the serpins were used to select a potentially exteriorly exposed and highly conserved region in the complexed form of α1-AT as a candidate ligand (carboxyl-terminal fragment, amino acids 359-374). We show here that synthetic peptides based on the sequence of this region bind specifically and anturably to human hepatoma cells and human monocytes (Kd = 4.0 × 10-8 M, 4.5 × 105 plasma membrane receptors per cell) and mediate increases in synthesis of α1-AT. Binding of peptide 105Y (Ser-Ile-Pro-Pro-Glu-Val-Lys-Phe-Asn-Lys-Pro-Phe-Val-Tyr-Leu-Ile) is blocked by α1-AT-elastase complexes, antithrombin III (AT Ill)-thrombin complexes, α1-antichymotrypsin (α1-ACT)-cathepsin G complexes, and, to a ser extent, complement component C1 inhibitor-C1s complexes, but not by the corresponding native proteins. Binding of peptide 105Y is also blocked by peptides with sequence corresponding to carboxyl-terminal fragments of the serpins AT III and α1-ACT, but not by peptides having the sequence of the extreme amino terminus of α1-AT. The results also show that peptide 105Y inhibits binding of 125I-labeled α1-AT-aducstase complexes. Thus, these studies demonstrate an abundant, relatively high-affinity cell surface receptor which recognizes serpin-enzyme complexes (SEC receptor). This receptor is capable of modulating the production of at least one of the serpins, α1-AT. Since the ligand specificity is similar to that previously described for in vivo clearance of serpin-enzyme complexes, the SEC receptor may also be involved in the clearance of certain serpin-enzyme complexes.

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