Structure of the Gene for Human Plasminogen Activator Inhibitor-2

Ye, Richard D.; Ahern, S; Beau, Michelle M. Le; Lebo, Roger V.; Sadler, J. Evan

doi:10.1016/s0021-9258(18)83572-4

Cited by 87 publications

(6 citation statements)

References 53 publications

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“…Ovalbumin represents the parent prototype of a unique family, within the serpin superfamily, that lack a typical cleavable signal sequence, but have been found to reside either intracellularly, extracellularly, or both (Remold-O'Donnell, 1993). The serpins previously classified as members of the ovalbumin branch are plasminogen activator inhibitor 2 (PAI-2)1 *** (Ye et al, 1989), an elastase inhibitor isolated from monocyte-like cells (El) (Dubin et al, 1992;Remold-O'Donnell et al, 1992), and a squamous cell carcinoma antigen (SCCA) (Suminami et al, 1991). Recently, we have isolated a 38-kDa serine proteinase inhibitor from the cytosolic fraction of a kidney epithelial cell line ( Morgenstern et al, 1993).…”

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confidence: 99%

Complementary DNA Cloning and Kinetic Characterization of a Novel Intracellular Serine Proteinase Inhibitor: Mechanism of Action with Trypsin and Factor Xa as Model Proteinases

Morgenstern¹,

Sprecher²,

Holth³

et al. 1994

Biochemistry

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The full-length cDNA encoding a novel human intracellular serine proteinase inhibitor has been sequenced and found to encode a 376 amino acid protein (M(r) approximately 42.5K) that we designate as cytoplasmic antiproteinase. Analysis of the primary structure revealed that the cytoplasmic antiproteinase has the majority of structural motifs conserved among the greater superfamily of serine proteinase inhibitors, or serpins. On the basis of several criteria such as amino acid identity and the absence of a classical N-terminal signal peptide, the cytoplasmic antiproteinase represents a new member of the intracellular serpin family. Further inspection of the cytoplasmic antiproteinase amino acid sequence identified three potential N-glycosylation sites and Arg341-Cys342 as the reactive site P1-P1' residues, respectively. We have also employed the slow binding kinetic approach to detail the mechanism of bovine trypsin and human factor Xa inhibition by the novel cytoplasmic antiproteinase. Inhibition of trypsin by the cytoplasmic antiproteinase was preceded by a two-step mechanism corresponding to the formation of an initial loose complex, followed by an isomerization step to a more stable, tight complex. The binding of the cytoplasmic antiproteinase to trypsin occurred with a second-order association rate constant of 2.8 x 10(6) M-1 s-1 and an overall equilibrium constant of 22.5 pM, demonstrating that the factor is a potent inhibitor of this proteinase. Under the appropriate conditions, the tight complex between trypsin and the cytoplasmic inhibitor was reversible, indicated by an exponential regeneration of proteinase amidolytic activity from the preformed complex. Therefore, the tight complex appears to be stabilized predominantly by reversible bonds that form between trypsin and the cytoplasmic inhibitor. In contrast to the inhibition of trypsin, the inhibition of factor Xa amidolytic activity by the cytoplasmic antiproteinase followed a single-step binding mechanism. The apparent first-order rate constant for factor Xa inhibition was found to increase as a linear function of the inhibitor concentration range studied. Formation of the inhibitory complex between factor Xa and the cytoplasmic antiproteinase occurred with a second-order association rate constant of approximately 1.3 x 10(5) M-1 s-1 and a equilibrium constant of 3.7 nM. These findings suggests that the cytoplasmic inhibitor may initially encounter significant energy barriers for proper alignment with the substrate binding cleft of factor Xa. However, once aligned, the reaction proceeds rapidly to a tight factor Xa.inhibitor complex that dissociates at a slow rate.

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confidence: 99%

Complementary DNA Cloning and Kinetic Characterization of a Novel Intracellular Serine Proteinase Inhibitor: Mechanism of Action with Trypsin and Factor Xa as Model Proteinases

Morgenstern¹,

Sprecher²,

Holth³

et al. 1994

Biochemistry

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“…Expression of PAI-2 has been detected in monocytes, macrophages, keratinocytes, fibroblasts, and the placenta [125,126]. The gene encoding human PAI-2, SERPINB2, is located on chromosome 18 (18q21-23) and consists of eight exons [127]. PAI-2 is a member of the serpin superfamily and was identified as a placental tissue-derived urokinase-type plasminogen activator (uPA) inhibitor and, to a lesser extent, a tPA inhibitor [126,[128][129][130].…”

Section: Pai-2mentioning

confidence: 99%

Fibrinogen and Antifibrinolytic Proteins: Interactions and Future Therapeutics

Pechlivani

Kearney

Ajjan

2021

IJMS

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Thrombus formation remains a major cause of morbidity and mortality worldwide. Current antiplatelet and anticoagulant therapies have been effective at reducing vascular events, but at the expense of increased bleeding risk. Targeting proteins that interact with fibrinogen and which are involved in hypofibrinolysis represents a more specific approach for the development of effective and safe therapeutic agents. The antifibrinolytic proteins alpha-2 antiplasmin (α2AP), thrombin activatable fibrinolysis inhibitor (TAFI), complement C3 and plasminogen activator inhibitor-2 (PAI-2), can be incorporated into the fibrin clot by FXIIIa and affect fibrinolysis by different mechanisms. Therefore, these antifibrinolytic proteins are attractive targets for the development of novel therapeutics, both for the modulation of thrombosis risk, but also for potentially improving clot instability in bleeding disorders. This review summarises the main properties of fibrinogen-bound antifibrinolytic proteins, their effect on clot lysis and association with thrombotic or bleeding conditions. The role of these proteins in therapeutic strategies targeting the fibrinolytic system for thrombotic diseases or bleeding disorders is also discussed.

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“…T he grouping of PA I2 and the ovalbum in related sequences is well supported and is perhaps surprising considering th at the divergence of the birds and m am m als is thought to have occurred ab o u t 180 million years ago. T he finding th at these sequences lack an am ino term inal signal sequence, having instead an internal sequence th at serves the same purpose (P alm iter et al 1978;Ye et al 1988Ye et al , 1989von Heijne et al 1991), gives support to their being related. It is likely th at ovalbum in has changed from being inhibitory to acting as storage protein.…”

Section: ( C) Gene Tre E a N A Ly Sismentioning

confidence: 99%

Evolutionary relationships among the serpins

Marshall

1993

Phil. Trans. R. Soc. Lond. B

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The serpins are a widely distributed group of serine proteinase inhibitors found in plants, birds, mammals and viruses. Despite the great evolutionary divergence of these organisms, their serpins are highly conserved, both in sequence and structurally. Amino acid sequences were aligned by a combination of automatic algorithms and by consideration of conserved structural elements in those serpins for which crystal structures exist. The program HOMED was used which allowed the alignment of amino acids to be simultaneously converted into the equivalently aligned nucleotide sequences. The aligned amino acids were used as the basis for superposition of the four known three-dimensional structures for which coordinates are available and compared with an optimal three-dimensional superposition in order to estimate the reliability of the sequence alignment. Phylogenetic relationships implied by these nucleotide sequence alignments were determined by the method of maximum parsimony. The proposed gene tree suggested that as much diversity existed between the plant serpin and mammalian serpins as was present among mammalian serpins and provided further evidence that the architecture of serpin molecules is highly constrained.

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Structure of the Gene for Human Plasminogen Activator Inhibitor-2

Cited by 87 publications

References 53 publications

Complementary DNA Cloning and Kinetic Characterization of a Novel Intracellular Serine Proteinase Inhibitor: Mechanism of Action with Trypsin and Factor Xa as Model Proteinases

Complementary DNA Cloning and Kinetic Characterization of a Novel Intracellular Serine Proteinase Inhibitor: Mechanism of Action with Trypsin and Factor Xa as Model Proteinases

Fibrinogen and Antifibrinolytic Proteins: Interactions and Future Therapeutics

Evolutionary relationships among the serpins

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