NDLI: Expression of a Novel Chimeric-Truncated tPA in Pichia pastoris with Improved Biochemical Properties

Content Provider	Springer Nature Link
Author	Saadatirad, Amirhossein Sardari, Soroush Kazemali, Mohammadreza Zarei, Najmeh Davami, Fatemeh Barkhordari, Farzaneh Adeli, Ahmad Mahboudi, Fereidoun
Copyright Year	2014
Abstract	Thrombolytic therapy by plasminogen activators (PAs) has been a main goal in the treatment of acute myocardial infarction. Despite improved outcomes of currently available thrombolytic therapies, all these agents have different drawbacks that may result in less than optimal outcomes. In order to make tissue plasminogen activator (tPA) more potent, while being more resistant to plasminogen activator inhibitor-1 (PAI-1) and having a higher affinity to fibrin, a new chimeric-truncated form of tPA (CT tPA) was designed and expressed in Pichia pastoris. This novel variant consists of a finger domain of Desmoteplase, an epidermal growth factor (EGF) domain, a kringle 1 (K1) domain, a kringle 2 (K2) domain, in which the lysine binding site (LBS) was deleted, and a protease domain, where the four amino acids lysine 296, arginine 298, arginine 299, and arginine 304 were substituted by aspartic acid. The chimera CT tPA showed 14-fold increase in its activity in the presence of fibrin compared to the absence of fibrin. Furthermore, CT tPA showed about 10-fold more potency than commercially available full-length tPA (Actylase$^{®}$) and provided 1.2-fold greater affinity to fibrin. A residual activity of only 68 % was observed after incubation of Actylase$^{®}$ with PAI-1, however, 91 % activity remained for CT tPA. These promising findings suggest that the novel CT tPA variant might be an acceptable PA with superior characteristics and properties.
Starting Page	1143
Ending Page	1150
Page Count	8
File Format	PDF
ISSN	10736085
Journal	Molecular Biotechnology
Volume Number	56
Issue Number	12
e-ISSN	15590305
Language	English
Publisher	Springer US
Publisher Date	2014-08-21
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Chimeric-truncated tPA Pichia pastoris Desmodous rotendus PA PAI-1 resistant Fibrin affinity Biotechnology Biochemistry Cell Biology Protein Science Biological Techniques Human Genetics
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biochemistry Bioengineering Applied Microbiology and Biotechnology Biotechnology

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Overexpression and Biochemical Characterization of a Thermostable Phytase from Bacillus subtilis US417 in Pichia pastoris

Secreted expression of truncated capsid protein from porcine circovirus type 2 in Pichia pastoris

Expression in Pichia pastoris and immunological evaluation of a truncated dengue envelope protein

Cloning, Heterologous Expression, Purification and Characterization of M12 Mutant of Aspergillus niger Glucose Oxidase in Yeast Pichia pastoris KM71H

Comparison of the Heterologous Expression of Trichoderma reesei Endoglucanase II and Cellobiohydrolase II in the Yeasts Pichia pastoris and Yarrowia lipolytica

A Thermolabile Aspartic Proteinase from Mucor mucedo DSM 809: Gene Identification, Cloning, and Functional Expression in Pichia pastoris

Streptomyces violaceoruber Phospholipase A2: Expression in Pichia pastoris, Properties, and Application in Oil Degumming

Expression of Recombinant Proteins in Pichia Pastoris

Codon Optimization of the “Bos Taurus Chymosin” Gene for the Production of Recombinant Chymosin in Pichia pastoris

Expression of a Novel Chimeric-Truncated tPA in Pichia pastoris with Improved Biochemical Properties

Similar Documents

Overexpression and Biochemical Characterization of a Thermostable Phytase from Bacillus subtilis US417 in Pichia pastoris

Secreted expression of truncated capsid protein from porcine circovirus type 2 in Pichia pastoris

Expression in Pichia pastoris and immunological evaluation of a truncated dengue envelope protein

Cloning, Heterologous Expression, Purification and Characterization of M12 Mutant of Aspergillus niger Glucose Oxidase in Yeast Pichia pastoris KM71H

Comparison of the Heterologous Expression of Trichoderma reesei Endoglucanase II and Cellobiohydrolase II in the Yeasts Pichia pastoris and Yarrowia lipolytica

A Thermolabile Aspartic Proteinase from Mucor mucedo DSM 809: Gene Identification, Cloning, and Functional Expression in Pichia pastoris

Streptomyces violaceoruber Phospholipase A2: Expression in Pichia pastoris, Properties, and Application in Oil Degumming

Expression of Recombinant Proteins in Pichia Pastoris

Codon Optimization of the “Bos Taurus Chymosin” Gene for the Production of Recombinant Chymosin in Pichia pastoris

Expression of a Novel Chimeric-Truncated tPA in Pichia pastoris with Improved Biochemical Properties