title: Exploring hemostatic and thrombolytic potential of heynein - A cysteine protease from Ervatamia heyneana latex.
abstract: BACKGROUND
  The latex of Ervatamia heyneana (Wall.) T. Cooke plant has been used for wound healing and various skin diseases by Indian tribes and folklore.
  
  
  OBJECTIVE
  To validate the scientific basis of heynein - a key protease of Ervatamia heyneana, in hemostasis and wound healing process.
  
  
  METHODS
  The latex from E. heyneana was processed and subjected to two step purification. The purified heynein was assayed for proteolytic activity using casein as substrate and also attested by zymography. The inhibition studies confirmed the nature of heynein. Pure fibrinogen was used for fibrinogenolytic activity and citrated plasma was used for coagulant and fibrinolytic activities. The edema inducing action and hemorrhagic activity of heynein were assessed on mice model.
  
  
  RESULTS
  The purified heynein exhibited proteolytic activity, which was confirmed by caseinolytic assay and zymography. The inhibition studies confirmed heynein to be a cysteine protease. Heynein showed complete hydrolysis of all the three subunits of human fibrinogen (Aα, Bβ, γ). It exhibited strong pro-coagulant activity by reducing plasma clotting time from 248 to 39s at 40µg concentration. Heynein cleaved α polymer subunit in fibrin clot and did not induce edema and hemorrhage in mice models. The non-hemorrhagic nature was supported with histopathological studies of skin samples.
  
  
  CONCLUSIONS
  Heynein displays strong pro-coagulant action associated with fibrin(ogen)olytic activity. This provides basis for the observed pharmacological action of Ervatamia heyneana and thereby justifies its use in folk medicine.
authors.affiliation: Department of Biotechnology, JSS Science and Technology University, JSS Technical Institutions Campus, Mysuru 570 006, Karnataka, India.
authors.forename: Priyanka
authors.initials: P
authors.lastname: Uday
authors.affiliation: Department of Biotechnology, JSS Science and Technology University, JSS Technical Institutions Campus, Mysuru 570 006, Karnataka, India.
authors.forename: M
authors.initials: M
authors.lastname: Maheshwari
authors.affiliation: Department of Studies in Biosciences, University of Mysore, Hemagangothri, Hassan, Karnataka 570006, India.
authors.forename: P
authors.initials: P
authors.lastname: Sharanappa
authors.affiliation: Department of Biotechnology, JSS Science and Technology University, JSS Technical Institutions Campus, Mysuru 570 006, Karnataka, India.
authors.forename: Zohara
authors.initials: Z
authors.lastname: Nafeesa
authors.affiliation: Department of Biotechnology, JSS Science and Technology University, JSS Technical Institutions Campus, Mysuru 570 006, Karnataka, India.
authors.forename: Vivek Hamse
authors.initials: VH
authors.lastname: Kameshwar
authors.affiliation: Department of Studies in Chemistry, University of Mysore, Mysuru 570 006, Karnataka, India.
authors.forename: B S
authors.initials: BS
authors.lastname: Priya
authors.affiliation: Department of Biotechnology, JSS Science and Technology University, JSS Technical Institutions Campus, Mysuru 570 006, Karnataka, India. Electronic address: nanjuchem@gmail.com.
authors.forename: S
authors.initials: S
authors.lastname: Nanjunda Swamy
mesh_terms: D000328:Adult; D000818:Animals; D029062:Apocynaceae; D057056:Cysteine Proteases / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D005340:Fibrinogen / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D006470:Hemorrhage / Q000188:drug therapy / Q000378:metabolism; D006487:Hemostasis / Q000187:drug effects / Q000502:physiology; D006490:Hemostatics / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D007840:Latex / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D008297:Male; D051379:Mice; D024301:Plant Bark; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D013927:Thrombosis / Q000188:drug therapy / Q000378:metabolism; D055815:Young Adult
chemical_list: D005343:Fibrinolytic Agents; D006490:Hemostatics; D007840:Latex; D010936:Plant Extracts; D005340:Fibrinogen; D057056:Cysteine Proteases
keywords: Ervatamia heyneana (Wall.) T. Cooke; Fibrino(geno)lytic; Hemostasis; Heynein (cysteine protease); Pro–coagulant