Identification of myxomaviral serpin reactive site loop sequences that regulate innate immune responses

Erbin Dai, Kasinath Viswanathan, Ming Sun Yun, Xing Li, Ying Liu Li, Babajide Togonu-Bickersteth, Jakob Richardson, Colin Macaulay, Piers Nash, Peter Turner, Steven H. Nazarian, Richard Moyer, Grant McFadden, Alexandra R. Lucas

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

The thrombolytic serine protease cascade is intricately involved in activation of innate immune responses. The urokinase-type plasminogen activator and receptor form complexes that aid inflammatory cell invasion at sites of arterial injury. Plasminogen activator inhibitor-1 is a mammalian serpin that binds and regulates the urokinase receptor complex. Serp-1, amyxomaviral serpin, also targets the urokinase receptor, displaying profound anti-inflammatory and anti-atherogenic activity in a wide range of animal models. Serp-1 reactive center site mutations, mimicking known mammalian and viral serpins, were constructed in order to define sequences responsible for regulation of inflammation. Thrombosis, inflammation, and plaque growth were assessed after treatment with Serp-1, Serp-1 chimeras, plasminogen activator inhibitor-1, or unrelated viral serpins in plasminogen activator inhibitor or urokinase receptor-deficient mouse aortic transplants. Altering the P1-P1′ Arg-Asn sequence compromised Serp-1 protease-inhibitory activity and anti-inflammatory activity in animal models; P1-P1′ Ala-Ala mutants were inactive, P1 Met increased remodeling, and P1′ Thr increased thrombosis. Substitution of Serp-1 P2-P7 with Ala6 allowed for inhibition of urokinase but lost plasmin inhibition, unexpectedly inducing a diametrically opposed, proinflammatory response with mononuclear cell activation, thrombosis, and aneurysm formation (p < 0.03). Other serpins did not reproduce Serp-1 activity; plasminogen activator inhibitor-1 increased thrombosis (p < 0.0001), and unrelated viral serpin, CrmA, increased inflammation. Deficiency of urokinase receptor in mouse transplants blocked Serp-1 and chimera activity, in some cases increasing inflammation. In summary, 1) Serp-1 anti-inflammatory activity is highly dependent upon the reactive center loop sequence, and 2) plasmin inhibition is central to anti-inflammatory activity.

Original languageEnglish (US)
Pages (from-to)8041-8050
Number of pages10
JournalJournal of Biological Chemistry
Volume281
Issue number12
DOIs
StatePublished - Mar 24 2006
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Dai, E., Viswanathan, K., Yun, M. S., Li, X., Li, Y. L., Togonu-Bickersteth, B., Richardson, J., Macaulay, C., Nash, P., Turner, P., Nazarian, S. H., Moyer, R., McFadden, G., & Lucas, A. R. (2006). Identification of myxomaviral serpin reactive site loop sequences that regulate innate immune responses. Journal of Biological Chemistry, 281(12), 8041-8050. https://doi.org/10.1074/jbc.M509454200