1rav

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1rav, resolution 2.2Å

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RECOMBINANT AVIDIN

Overview

The mature hen avidin encoded by a synthetic cDNA was expressed in, Escherichia coli in an insoluble form. After resolubilization, renaturation and purification, a recovery of about 20 mg/l cell culture, was obtained. ELISA assays indicated no apparent differences in biotin, binding between the natural and recombinant avidins. In addition, an, acidic avidin mutant, bearing the substitutions Lys3-->Glu, Lys9--> Glu, Arg26-->Asp and Arg124-->Leu of four exposed basic residues, was produced., The protein, expressed and renatured as wild-type avidin, showed unaltered, biotin-binding activity. The acidic pI (approximately 5.5) and lack of, aggregation of the mutant allowed easy electrophoretic analysis under, non-denaturing conditions of the protein alone and of its complexes with, ... [(full description)]

About this Structure

1RAV is a [Single protein] structure of sequence from [Gallus gallus]. Structure known Active Site: . Full crystallographic information is available from [OCA].

Reference

Biochemical characterization and crystal structure of a recombinant hen avidin and its acidic mutant expressed in Escherichia coli., Nardone E, Rosano C, Santambrogio P, Curnis F, Corti A, Magni F, Siccardi AG, Paganelli G, Losso R, Apreda B, Bolognesi M, Sidoli A, Arosio P, Eur J Biochem. 1998 Sep 1;256(2):453-60. PMID:9760187

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