Study of urinary proteomes in Anderson-Fabry disease

Figures & data

Table 1.  Clinical data of Anderson-Fabry disease patients

Patient number	Patient age	Gender	Mutation GAL, genotype	Mainz score	Alphagal leukocyte (nmol/mg h)	ERT	Therapy from	Drug	DM	ACE inhib., AT block.	S_creatinin (μmol/L)	U_creatinin(mmol/L)	HD	Transplantation	Proteinuria (g/L)
1	57	F	N215S	19	30.3	N			N	Y	53	11.5	N	N	<0.07
2	60	F	N215S	20	27.8	N			N	N	76	8.40	N	N	<0.07
3	69	F	c.674del59	27	23.6	Y	12/2001	Fabrazyme	N	Y	77	11.10	N	N	0.14
4	39	F	R342Q	14	28.6	N			N	N	78	1.6	N	N	<0.07
5	60	F	R342Q	15	8.5	Y	11/2004	Replagal	N	Y	85	18.3	N	N	0.17
6	46	F	delEx2	19	25.4	N			N	N	78	13.7	N	N	<0.07
7	55	F	R342Q	7	17.9	Y	12/204	Replagal	N	Y	57	18.6	N	N	0.13
8	64	F	I317T	38	22.3	N			N	Y	73	6.8	N	N	0.08
9	61	F	R342Q	54	21	N			Y	Y	93	4.3	N	N	0.09
10	40	M	c.674del59	38	0.7	Y	12/2004	Replagal	N	N	75	5.6	N	N (nephrectomy2006)	0.18
11	32	M	c.674del59	14	0.7	Y	12/2003	Replagal	N	N	67	6.9	N	N	0.08
12	43	M	c.674del59	31	0.3	Y	12/2003	Fabrazyme	N	N	82	9.5	N	N	0.1
13	42	M	delEx2	22	2.5	Y	1/2004	Fabrazyme	N	N	73	10.5	N	N	0.14
14	40	M	R342Q	25	1.6	Y	2/2004	Fabrazyme	N	N	75	7.7	N	N	0.08
15	18	M	c.674del59	16	0,2	N	2007	Fabrazyme	N	N	69	3,3	N	N	<0.07
16	52	M	delEx2	34	0.22	Y	1/2004	Fabrazyme	N	Y	71	9.5	N	N	0.24
17	59	M	c.674del59	51	2.6	Y	7/2001	Fabrazyme	N	N	185	2.4	N	N	0.37
18	55	M	L294X	32	0.4	Y	11/2004	Fabrazyme	N	Y	645	6.3	Y	Y (1984, HD started from 2005)	0.61
19	51	M	R342Q	47	1.0	Y	11/2004	Fabrazyme	N	Y	218	6.1	Y	Y (HD started in 2001, next transpl. 2006)	0.38
20	48	M	c.674del59	34	1.82	Y	2/2004	Fabrazyme	N	Y	122	6.2	N	N	0.66

Note: DM, diabetes mellitus; ERT, enzyme replacement therapy; HD, hemodialysis; ACE, inhibitors of angiotensin-converting enzyme; AT, blocker of angiotensin receptor; Y, yes; N, no; F, female; M, male.

Figure 1. Comparison of 2D electrophoreograms of urine samples collected from patients with Fabry disease (A, B) and healthy controls (C, D). The urine samples were analyzed immediately after their collection and short-time centrifugation, using 7 cm polyacrylamide strips, pH 3–10 L, and SDS-PAGE. BenchMark^TM Protein Ladder (Invitrogen, Glasgow, UK) was used as a molecular mass marker.

Table 2.  Changes of selected proteins observed using Phoretix 2D expression software version 2005

No.	Protein	Anderson-Fabry disease (average ± SD) %^a	Expected molecular weight (kDa)	Expected pI
1	Prostaglandin H2 d-isomerase	398 ± 25	28	6.1
2	Complement-c1q tumor necrosis factor-related protein	289 ± 17	23	5.8
3	Ig kappa chain V–III	591 ± 7	27	5.5

Note: *p < 0.05 versus healthy controls. Samples were collected from 20 different patients with Anderson-Fabry disease. ^aN = 20: [(amount of protein in the sample with Anderson-Fabry disease/amount of protein in healthy control) × 100].

Figure 2. Changes in the protein amount in the samples collected from 20 patients with Fabry disease compared with those in healthy controls. *p < 0.05 versus healthy controls.

Figure 3. A representative 2D gel image of the Fabry disease proteome, depicting all identified proteins. All urine samples were analyzed immediately after their collection using 7 cm polyacrylamide strips, pH 3–10 L, and SDS-PAGE.

Table 3.  Comparison of experimental and theoretical molecular size and isoelectric point of identified proteins by MALDI-TOF MS

No.	Protein	Experimental pI	Theoretical pI	Experimental molecular weight	Theoretical molecular weight
1	Alpha-1-antitrypsin	5.1	5.4	48	47
2	Alpha-1-antitrypsin	4.9	5.4	47	47
3	Alpha-1-microglobulin/bikunin precursor	5.2	5.9	40	40
4	Alpha-1-microglobulin/bikunin precursor	5.1	5.9	40	40
5	Ig kappa chain V–III	5.5	5.8	27	23
6	Complement-c1q tumor necrosis factor-related protein	5.8	6.1	23	25
7	Prostaglandin H2 d-isomerase	6.1	7.7	28	21

Note: *p < 0.05 experimental versus theoretical parameter, N = 20.