Arginine hydrochloride enhances chaperone-like activity of alpha crystallin
First Claim
1. A method for enhancing molecular chaperone activity of α
- -crystalline (comprising of forms α
-crystalline and α
B-crystalline) with a biological compatible amino acid molecule of Arginine Hydrochloride, said method comprising the steps of;
(a) isolating and purifying α
-crystalline from calf eye lenses by convention methods (as described in reference
24), and (b) reacting α
-crystalline in the presence of phosphate buffer of pH 7.4 with Arg.HCl and insulin or ξ
-crystalline in presence or absence of DTT, and (c) observing the enhancement in chaperone activity of α
-crystalline in presence of Arg.HCl by fluorescence spectrophotometer.
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Abstract
The present invention provides that arginine, a biologically compatible molecule that is known to bind to the peptide backbone and negatively charged side-chains, increases the chaperone-like activity of a-crystallin significantly. Arginine, interestingly, restores the activity of mutant protein to a considerable extent. The invention shows the effect of arginine on the structural changes of a-crystallin by circular dichroism, fluorescence and glycerol gradient sedimentation. The invention shows that arginine brings about subtle changes in the tertiary structure and significant changes in the quaternary structure of a-crystallin and enhances its chaperone activity significantly. This invention thus has role in designing strategies to improve chaperone function for therapeutic applications.
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Citations
18 Claims
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1. A method for enhancing molecular chaperone activity of α
- -crystalline (comprising of forms α
-crystalline and α
B-crystalline) with a biological compatible amino acid molecule of Arginine Hydrochloride, said method comprising the steps of;
(a) isolating and purifying α
-crystalline from calf eye lenses by convention methods (as described in reference
24), and(b) reacting α
-crystalline in the presence of phosphate buffer of pH 7.4 with Arg.HCl and insulin or ξ
-crystalline in presence or absence of DTT, and(c) observing the enhancement in chaperone activity of α
-crystalline in presence of Arg.HCl by fluorescence spectrophotometer. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
- -crystalline (comprising of forms α
Specification