title: Molecular requirements for safer generation of thrombolytics by bioengineering the tissue-type plasminogen activator A chain.
abstract: BACKGROUND
  Thrombolysis with tissue-type plasminogen activator (t-PA) is the only treatment approved for acute ischemic stroke. Although t-PA is an efficient clot lysis enzyme, it also causes damage to the neurovascular unit, including hemorrhagic transformations and neurotoxicity.
  
  
  OBJECTIVE
  On the basis of the mechanism of action of t-PA on neurotoxicity, we aimed at studying the molecular requirements to generate safer thrombolytics.
  
  
  METHODS
  We produced original t-PA-related mutants, including a non-cleavable single-chain form with restored zymogenicity (sc*-t-PA) and a t-PA modified in the kringle 2 lysine-binding site (K2*-t-PA). Both sc*-t-PA and K2*-t-PA showed fibrinolytic activities similar to that of wild-type t-PA on both euglobulin-containing and plasma-containing clots. In contrast to wild-type t-PA, the two mutants did not promote N-methyl-d-aspartate receptor-mediated neurotoxicity.
  
  
  CONCLUSIONS
  We designed t-PA mutants with molecular properties that, in contrast to t-PA, do not induce neurotoxicity.
authors.affiliation: Inserm, Inserm UMR-S U919, University of Caen Basse-Normandie, Serine Proteases and Pathophysiology of Neurovascular Unit, GIP Cyceron, Caen, France.
authors.forename: J
authors.initials: J
authors.lastname: Parcq
authors.forename: T
authors.initials: T
authors.lastname: Bertrand
authors.forename: A F
authors.initials: AF
authors.lastname: Baron
authors.forename: Y
authors.initials: Y
authors.lastname: Hommet
authors.forename: E
authors.initials: E
authors.lastname: Anglès-Cano
authors.forename: D
authors.initials: D
authors.lastname: Vivien
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D057005:Bioengineering / Q000295:instrumentation / Q000379:methods; D019149:Bioreactors; D016923:Cell Death / Q000187:drug effects; D015195:Drug Design; D005342:Fibrinolysis / Q000187:drug effects*; D005343:Fibrinolytic Agents / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D057809:HEK293 Cells; D006801:Humans; D018082:Kringles; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D020258:Neurotoxicity Syndromes / Q000209:etiology / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D017354:Point Mutation; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D013329:Structure-Activity Relationship; D015912:Thrombolytic Therapy / Q000009:adverse effects; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology* / Q000633:toxicity; D014162:Transfection
chemical_list: D005343:Fibrinolytic Agents; D016194:Receptors, N-Methyl-D-Aspartate; D011994:Recombinant Proteins; D010959:Tissue Plasminogen Activator