TABLE 1

Effect of amino acid at position 87 in CYP102A1 M11H on the GSH conjugation to reactive metabolites formed by oxidative bioactivation of clozapine

Concentration of metabolites (μM) formed after 30-min incubations of 200 nM CYP102A1 with 500 μM clozapine and 8 μM GST P1-1 in the presence of 100 μM GSH. Values represent averages of three individual experiments; S.E.s were always less than 10%.

	C-1	C-2	C-3	C-4	C-5	CG-1	CG-2	CG-3	CG-4	CG-5	CG-6	CG-7	CG-8	Specific Activity^{^a}	Percentage Conversion
m/z (MH⁺)	343	313	301	287	329	632	618	632	632	632	598	903	584
Apolar side chain
Gly87	0.4	0.7	0.1	N.D.	N.D.	1.1	N.D.	0.1	N.D.	1.2	2.7	0.5	0.4	1.2	1.4
Ala87	35.1	82.5	1.1	2.0	7.8	13.7	0.7	1.0	0.3	14.4	26.1	4.0	2.5	31.9	38.2
Val87	15.1	90.1	0.9	1.0	7.1	9.3	0.7	0.8	0.3	11.7	15.8	5.5	2.9	26.9	32.3
Leu87	1.0	9.0	0.3	0.1	N.D.	3.2	0.1	0.5	0.3	4.7	6.3	1.2	0.6	4.6	5.5
Ile87	6.8	71.4	0.5	0.3	0.6	9.6	0.5	0.8	0.4	N.D.	20.7	2.9	1.6	21.6	25.9
Phe87	8.7	10.5	0.2	0.2	N.D.	18.2	0.2	0.9	0.3	16.7	37.2	5.3	2.2	16.8	20.1
Trp87	11.6	46.9	N.D.	N.D.	N.D.	5.0	N.D.	N.D.	N.D.	5.4	7.6	2.7	0.8	13.3	16.0
Met87	0.7	5.5	0.1	0.1	N.D.	2.8	N.D.	0.3	N.D.	3.5	5.1	N.D.	0.7	3.2	3.8
Pro87	1.4	4.6	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.Q.^{^b}	1.2
Polar uncharged side chain
Tyr87	16.5	73.4	0.9	0.9	5.2	6.7	0.3	0.5	0.2	8.1	11.5	3.0	1.1	21.4	25.7
Gln87	3.2	6.4	0.1	0.1	N.D.	8.5	0.1	0.7	0.3	9.7	13.9	1.9	2.1	7.9	9.4
Asn87	0.7	3.2	0.2	0.2	N.D.	2.3	N.D.	0.2	N.D.	2.8	4.7	1.0	0.7	2.6	3.2
Cys87	0.2	1.0	0.3	N.D.	N.D.	1.0	N.D.	N.D.	N.D.	1.0	2.0	N.D.	N.D.	0.9	1.1
Ser87	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.
Thr87	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.
Polar charged side chain
Asp87	8.5	13.6	N.D.	N.D.	N.D.	3.7	N.D.	N.D.	N.D.	3.7	7.1	0.3	0.3	N.Q.^{^b}	7.4
Glu87	0.6	4.3	0.1	0.1	N.D.	1.7	N.D.	N.D.	N.D.	1.7	3.1	0.5	N.D.	2.0	2.4
His87	0.9	1.3	0.3	0.1	N.D.	0.9	N.D.	N.D.	N.D.	1.1	1.6	N.D.	N.D.	1.0	1.2
Arg87	1.7	4.5	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.	1.0	1.2

N.D., not detected.
↵a Presented as nmol product/nmol CYP102A1/30 min.
↵b N.Q., not quantified because P450 concentration could not be quantified (CO difference spectrum only showed a peak at 420 nm).