Comparison of complexes formed between crustacean and vertebrate trypsins with bovine pancreatic trypsin inhibitor : a key to reach extreme stability?

Molnár, Tamás and Vörös, Judit and Szeder, Bálint and Takáts, Kornél and Kardos, József and Katona, Gergely and Gráf, László (2013) Comparison of complexes formed between crustacean and vertebrate trypsins with bovine pancreatic trypsin inhibitor : a key to reach extreme stability? FEBS Journal, 280 (22). pp. 5750-5763. ISSN 1742-464X (Unpublished)

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Abstract

This paper provides evidence for the extremely high resistance of a complex of crayfish trypsin (CFT) and bovine pancreatic trypsin inhibitor (BPTI) against heating and chemical denaturing agents such as sodium dodecyl sulfate (SDS) and urea. To dissociate this complex 15 minute boiling in SDS was necessary as compared to a complex of bovine trypsin and BPTI that dissociates in SDS without boiling. The CFT-BPTI complex remained stable even in 9 M urea, while the bovine trypsin-BPTI complex started to dissociate at about 4 M urea. The melting temperature of the bovine trypsin-BPTI and CFT-BPTI complexes, as determined by differential scanning calorimetry, were found to be 79.6 and 100.1 oC, respectively. The behaviour of the apo-enzymes did not provide a definite clue to the differential effects of BPTI on their stabilities, although CFT was found to have a less stable structure as compared to that of bovine trypsin and in the X-ray structure of the CFT-BPTI complex we identified a few extra binding sites for BPTI. Though the structural cause of the extreme stability of the CFT-BPTI complex is not yet fully understood, our study could be a starting point to develop new protein complexes with enhanced stability.

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