Characterization of cold-adapted Atlantic cod (Gadus morhua) trypsin I--kinetic parameters, autolysis and thermal stability. 2010

Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
Department of Biochemistry, Science Institute University of Iceland, Dunhaga 3, 107 Reykjavík, Iceland.

Atlantic cod trypsin I is a highly active cold-adapted protease. This study aimed at further characterization of this enzyme with respect to kinetic parameters, sites of autolysis and stability. For that purpose, trypsin I was purified by anion exchange chromatography. Its purity and identity was verified by SDS-PAGE analysis and mass spectrometry. Concomitantly, another cod trypsin isozyme, trypsin X, previously only described from its cDNA sequence was detected in a separate peak from the ion exchange chromatogram. There was a stepwise increase in the catalytic efficiency (k(cat)/K(m)) of cod trypsin I obtained with substrates containing one to three amino acid residues. As expected, the activity of trypsin I was maintained for longer periods of time at 15 degrees C than at higher temperatures. The residues of the trypsin I molecule most sensitive to autolysis were identified using Edman degradation. Eleven autolytic cleavage sites were detected within the trypsin I molecule. Unfolding experiments demonstrated that autolysis is a contributing factor in the stability of trypsin I. In addition, the data shows that cod trypsin I is less stable towards thermal unfolding than its mesophilic bovine analogue.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D002417 Cattle Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor. Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D003080 Cold Temperature An absence of warmth or heat or a temperature notably below an accustomed norm. Cold,Cold Temperatures,Temperature, Cold,Temperatures, Cold
D004791 Enzyme Inhibitors Compounds or agents that combine with an enzyme in such a manner as to prevent the normal substrate-enzyme combination and the catalytic reaction. Enzyme Inhibitor,Inhibitor, Enzyme,Inhibitors, Enzyme
D004795 Enzyme Stability The extent to which an enzyme retains its structural conformation or its activity when subjected to storage, isolation, and purification or various other physical or chemical manipulations, including proteolytic enzymes and heat. Enzyme Stabilities,Stabilities, Enzyme,Stability, Enzyme
D000222 Adaptation, Physiological The non-genetic biological changes of an organism in response to challenges in its ENVIRONMENT. Adaptation, Physiologic,Adaptations, Physiologic,Adaptations, Physiological,Adaptive Plasticity,Phenotypic Plasticity,Physiological Adaptation,Physiologic Adaptation,Physiologic Adaptations,Physiological Adaptations,Plasticity, Adaptive,Plasticity, Phenotypic
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001665 Binding Sites The parts of a macromolecule that directly participate in its specific combination with another molecule. Combining Site,Binding Site,Combining Sites,Site, Binding,Site, Combining,Sites, Binding,Sites, Combining
D013050 Spectrometry, Fluorescence Measurement of the intensity and quality of fluorescence. Fluorescence Spectrophotometry,Fluorescence Spectroscopy,Spectrofluorometry,Fluorescence Spectrometry,Spectrophotometry, Fluorescence,Spectroscopy, Fluorescence
D014357 Trypsin A serine endopeptidase that is formed from TRYPSINOGEN in the pancreas. It is converted into its active form by ENTEROPEPTIDASE in the small intestine. It catalyzes hydrolysis of the carboxyl group of either arginine or lysine. EC 3.4.21.4. Tripcellim,Trypure,beta-Trypsin,beta Trypsin

Related Publications

Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
January 2017, Biochimica et biophysica acta. Proteins and proteomics,
Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
November 2000, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology,
Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
January 2006, Biochimica et biophysica acta,
Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
March 2019, International journal of biological macromolecules,
Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
November 2010, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP,
Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
January 2012, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc,
Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
August 1971, Canadian journal of biochemistry,
Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
January 2004, Protein expression and purification,
Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
December 2010, Fish & shellfish immunology,
Bjarki Stefansson, and Linda Helgadóttir, and Sigridur Olafsdottir, and Agústa Gudmundsdottir, and Jón B Bjarnason
January 1990, Comparative biochemistry and physiology. B, Comparative biochemistry,
Copied contents to your clipboard!