Selective stabilization of a labile mutant form of bovine pancreatic ribonuclease A by antibodies |
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Authors: | H Younus J Köditz M Saleemuddin R Ulbrich-Hofmann |
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Institution: | (1) Interdisciplinary Biotechnology Unit and Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, 202002, India;(2) Department of Biochemistry/Biotechnology, Martin Luther University Halle-Wittenberg, D-06099 Halle, Germany |
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Abstract: | The region between the amino acids 31-46 was previously identified as being first exposed during thermal unfolding of bovine pancreatic ribonuclease A (RNase). The exchange of one amino acid (Leu35toSer) in this unfolded region of RNase is shown to have a dramatic destabilizing effect ( Tm=9 °C). Antibodies raised against a peptide corresponding to the sequence of the labile region, S32-V43, of RNase were effective in stabilizing L35S-RNase against thermal inactivation (65 °C for 2 h) and surpassed the stabilization effect of antiRNase antibodies. An 11% contribution to the stabilizing effect of antiRNase antibodies resulted from antibodies recognizing the unfolding region of the enzyme. |
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Keywords: | antibodies enzyme stabilization mutants ribonuclease A unfolding region |
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