Abstract
The design of chimeric proteins is a major field of interest in structural biology and biotechnology. The successful design of the chimeric protein composed by the minimized reactive site domain of the low-molecular-mass trypsin inhibitor from Brassica napus (var. oleifera) seed (Ser3-Lys35; mini-RTI-III) and murine dihydrofolate reductase (DHFR) is reported here. The DHFR-mini-RTI-III chimeric protein was expressed in Escherichia coli, purified by metal-chelate affinity chromatography and oxidatively refolded. The affinity of the purified and refolded DHFR-mini-RTI-III for bovine trypsin (K = 5.0 x 10-10 M) was closely similar to that determined for native RTI-III (K = 2.9 x 10-10 M), at pH 8.2 and 22.0°C. DHFR-mini-RTI-III may be regarded as a tool in structure-function studies and for developing multifunctional and multidomain proteinase inhibitors. (C) 2000 Academic Press.
Original language | English |
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Pages (from-to) | 817-820 |
Number of pages | 4 |
Journal | Biochemical and Biophysical Research Communications |
Volume | 275 |
Issue number | 3 |
DOIs | |
Publication status | Published - Sep 7 2000 |
Keywords
- Chimeric oil rape mini-trypsin-inhibitor
- E. coli expression
- Inhibitory properties
- Oil rape (Brassica napus var. oleifera) trypsin inhibitor
- Oxidative refolding
- White mustard (Sinapis alba L.) trypsin inhibitor
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology