Prolyl Endopeptidase Probes
AssigneeSRI International
InventorsAmit K. Galande, Douglas Stuart Watson, Krishna Kodukula, Kalyani Jambunathan
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Abstract
Prolyl endopeptidase (PE) activity in lung samples is detected by contacting the lung sample with a probe comprising a —P—X— (or —X—P—, —P—X—P—) PE recognition site, wherein P is a prolyl bioisostere, X is a residue that is not a prolyl bioisostere or is a prolyl bioisostere flanked on each side by a residue that is not a prolyl bioisostere, and “-” is an amide bond, under conditions wherein PE activity of the sample specifically hydrolyzes an amide bond of the recognition site to generate an optical signal; and (b) detecting the signal.
Claims20
A method of detecting prolyl endopeptidase (PE) activity in a lung sample, comprising:
- (a) contacting the sample with a probe comprising a —P—X— or -P—X—P— PE recognition site, wherein P is a prolyl bioisostere, X is a residue that is not a prolyl bioisostere or is a prolyl bioisostere flanked on each side by a residue that is not a prolyl bioisostere, and “-” is an amide bond, under conditions wherein PE activity of the sample specifically hydrolyzes an amide bond of the recognition site to generate an optical signal;
- and
- (b) detecting the signal.
A method of detecting prolyl endopeptidase (PE) activity in each of a plurality of biological or physiological samples, comprising:
- (a) contacting each sample with a panel of different probes each comprising a —P—X— or —P—X—P— PE recognition site, wherein P is a prolyl bioisostere, X is a residue that is not a prolyl bioisostere or is a prolyl bioisostere flanked on each side by a residue that is not a prolyl bioisostere, and “-” is an amide bond, under conditions wherein PE activity of the sample specifically hydrolyzes an amide bond of the recognition site to generate an optical signal;
- and
- (b) detecting and comparing the resultant signals from each sample to determine differences in PE activity between the samples.
An internally quenched fluorogenic probe (IQFP) for prolyl endopeptidase (PE) activity comprising a —P—X— or -P—X—P— PE recognition site, wherein P is a prolyl bioisostere, X is a residue that is not a prolyl bioisostere or is a prolyl bioisostere flanked on each side by a residue that is not a prolyl bioisostere, and “-” is an amide bond, wherein one end of the recognition site is operably-linked to a first chromophore, and the other end of the recognition site is operably-linked to a second chromophore, wherein PE hydrolysis of an amide bond of the recognition generates an optical signal.
Dependencies are inferred from the claim text.