Isolation and analysation of soybean agglutinin-specific binding proteins for erythrocyte membrane in different animal species

Figures & data

Figure 1. Comparison of extraction effect of different lysis buffers on erythrocyte membrane in sodium salt (sds)-polyacrylamide gel electrophoresis (SDS-PAGE). M. Standard protein maker 1. Octylphenoxypolyethoxyethanol (NP-40) lysis buffer 2. Radio immunoprecipitation assay (RIPA) lysis buffer (medium) 3. RIPA lysis buffer (weak).

Figure 2. SDS-PAGE of erythrocyte membrane (EM) proteins in rabbit and pig. M. Standard protein maker 1. EM proteins of pig 2. EM proteins of rabbit.

Table 1. The soybean agglutinin specific binding cytoskeleton protein of EMs in three animal species.

		Rabbit					Swine					Bovine
No.	Protein name	AN	S	MW (kDa)	CP (%)	PI	AN	S	MW (kDa)	CP (%)	PI	AN	S	MW/kDa	CP (%)	PI
1	Keratin 10	G1T1V0	884	58,260	25.8	4.98	I3LDS3	597	58243	17.1	4.93	A6QNZ7	649	54987	16.2	5.01
2	Keratin 1	G1U9I8	609	64,422	14.4	8.6	F1SGG3	456	65381	12.2	8.18	G3N0V2	549	63341	13	8.41
3	Spectrin beta	G1SD01	48	268,766	1	5.38	F1SA60	530	271142	20.1	5.23	F1MKE9	591	269692	54.2	5.16
4	Spectrin alpha	G1STR7	48	281,789	3.1	4.94	F1RNB8	550	27767	26.6	4.92	F1MMQ8	650	282352	79.3	4.84
5	Keratin 5	G1TDN6	493	62,414	17.2	7.62	F1SGG6	250	63,668	18.7	7.74	M0QVZ6	608	60,800	25	7.63
6	Actin	P29751	166	42,071	21.1	5.3	I3LVD5	240	42,108	11.5	5.31	F1MRD0	105	42,167	16.5	5.16
7	Keratin 75	G1SHY2	335	61,390	12.5	8.43	F1SGI7	183	65,354	9	9.27	Q08D91	276	59,399	10.3	7.56
8	Keratin 14	G1T1Y7	324	48,662	15.1	4.87	F1S0L1	352	51,811	19.8	4.94	F1MC11	466	52,164	28.7	5.08
9	Keratin 3	G1SY72	296	64,858	10.2	7.6	I3LDM6	137	65,497	11.4	8.03	G3MXL3	459	63,082	14	5.78
10	Ankyrin 1	G1TG85	267	203,233	8	6.11	F1SE30	351	207,769	16.3	6	F1MY81	581	123,754	15.9	6.92
11	Keratin 8	G1SWS0	59	51,483	3.6	4.9	F1SGG2	63	54,394	7.4	5.7	P05786	141	53,594	11.7	5.6
12	Keratin 18	G1TMP1	137	48,347	5.8	5.33	F1SGG1	100	49,430	3.6	5.39	A6H7D3	68	49,218	3.6	5.33
13	Keratin 13	G1T4S1	454	48,712	19	4.86	F1S0K1	225	49,505	9.7	4.88	_	_	_	_	_
14	Ankyrin 2	G1SZP4	55	42,7545	0.6	4.98	F1S146	141	428,715	4.4	4.93	_	_	_	_	_
15	Keratin 35	G1T4P5	65	51,439	6.2	4.87	F1S0K7	99	51,637	3.5	4.95	_	_	_	_	_
16	Keratin 16	G1T4R6	332	49,878	12	49878	_	_	_	_	_	_	_	_	_	_
17	Keratin 20	G1TE74	147	84,376	4.6	4.89	_	_	_	_	_	_	_	_	_	_
18	Tubulin alpha chain	G1SL10	97	50,834	18.2	4.94	_	_	_	_	_	_	_	_	_	_
19	Keratin 222	G1U0K9	68	34,270	5.8	5.83	_	_	_	_	_	_	_	_	_	_
20	Keratin 7 C	G1TP66	53	55,448	7.8	5.8	_	_	_	_	_	_	_	_	_	_
21	Keratin 2	G1SUH8	618	63,646	19.1	8.98	_	_	_	_	_	E1B991	246	64,570	6.8	8.92
22	Keratin 7	G1SHZ4	239	40,889	11.5	4.88	_	_	_	_	_	Q29S21	288	51,603	10.3	5.79
23	Keratin 85	G1SHW8	40	56,850	5.1	7.03	_	_	_	_	_	Q0VD04	61	57,171	8.7	6.15
24	Keratin 15	_	_	_	_	_	F1S0K2	246	49,501	12	4.87	Q17QL7	295	49,083	12.6	4.83
25	Keratin 28	_	_	_	_	_	F1RXG2	180	50,560	5.4	5.08	Q148H6	160	51,314	7.3	5.19
26	Keratin 6 A	_	_	_	_	_	LOC100737483	172	60,375	15	8.53	M0QVY0	525	61,166	19.6	8.28
27	Keratin 19	_	_	_	_	_	F1S0J8	159	44,186	11.4	5.05	P08728	160	43,858	13.5	4.92
28	Keratin 34	_	_	_	_	_	F1S0K4	98	47,282	5.7	4.94	F1MSA6	197	45,782	19.4	4.86
29	Keratin 2 A	_	_	_	_	_	A5A759	259	66,510	8.3	7.47	_	_	_	_	_
30	Keratin 77	_	_	_	_	_	I3LNT6	202	62,707	8.3	5.89	_	_	_	_	_
31	Plakoglobin	_	_	_	_	_	Q8WNW3	108	82,539	2.4	5.75	_	_	_	_	_
32	Protein 4.2	_	_	_	_	_	F1SI48	106	77,185	3.6	6.93	_	_	_	_	_
33	Keratin 4	_	_	_	_	_	I3LQN8	62	57,648	6.7	7.45	_	_	_	_	_
34	Keratin 6 C	_	_	_	_	_	_	_	_	_	_	F1MUY2	460	61,275	19.4	8.58
35	Band 3 protein	_	_	_	_	_	_	_	_	_	_	Q9TUQ0	346	95,867	18.7	6
36	Keratin 17	_	_	_	_	_	_	_	_	_	_	A1L595	280	48,967	18.8	5.08
37	Keratin 24	_	_	_	_	_	_	_	_	_	_	F1MFW9	232	50,904	6.9	4.68
38	Keratin 28	_	_	_	_	_	_	_	_	_	_	Q148H6	183	51,314	7.3	5.19
39	Keratin 31	_	_	_	_	_	_	_	_	_	_	Q148I8	169	48,526	12.3	4.79
40	Protein 4.1	_	_	_	_	_	_	_	_	_	_	F1MCU7	158	58,782	13.3	6.45
41	Keratin 12	_	_	_	_	_	_	_	_	_	_	F1MJ66	152	50,303	7.9	4.55
42	Keratin 33B	_	_	_	_	_	_	_	_	_	_	A3KN26	90	44,440	10.7	4.81
43	Keratin 73	_	_	_	_	_	_	_	_	_	_	A7YWK3	88	59,540	3.5	6.96
44	Keratin 71	_	_	_	_	_	_	_	_	_	_	Q148H5	70	57,885	4	6.27
45	Keratin 86	_	_	_	_	_	_	_	_	_	_	E1B898	62	54,951	8.6	5.49
46	Keratin 83	_	_	_	_	_	_	_	_	_	_	A4FUZ0	62	55,665	8.5	5.43
47	Keratin 81	_	–	_	_	_	_	_	_	_	_	Q148H4	62	56,403	8.4	5.62
48	Vimentin	_	_	_	_	_	_	_	_	_	_	P48616	29	53,752	3.4	5.06

Accession number in the NCBInr database.

AC: accession number; MW: molecular weight; IP: isoelectric points; CP: coverage percentage; S: score.

Peptide score is shown as −10log(p), where p is the probability that the observed match is a random event. MASCOT protein scores (combined MS and MS/MS spectra) > 22 in rabbit, protein score >23 in pig, and >24 in bovine, were considered statistically significant (p < .05).

Table 2. The soybean agglutinin specific binding catalytic enzyme proteins in the EMs of three different animal species.

		Rabbit					Swine					Bovine
No.	Protein name	AN	S	MW (kDa)	CP (%)	PI	AN	S	MW (kDa)	CP (%)	PI	AN	S	MW/kDa	CP (%)	PI
1	Protein arginine methyltransferase 5	G1SGM2	72	73286	7.4	5.95	C3RZ98	26	73,252	4.4	5.88	F1N443	84	73,283	7.4	5.88
2	Methyltransferase like 3	G1SD33	28	65,012	2.9	6.05	F1S8J8	28	65,501	4.8	6.12		_	_	_	_
3	Pyruvate kinase	P11974	90	58,524	6	7.62	_	_	_	_	_	_	_	_	_	_
4	Enolase 2	G1SK04	26	47,536	12.7	4.95	_	_	_	_	_	_	_	_	_	_
5	Glutathione S-transferase theta-4	G1SN46	23	28,140	2.5	6.76	_	_	_	_	_	_	_	_	_	_
6	Trypsinogen	_	_	_	_	_	F1SRS2	306	26,563	7.3	6.85		_	_	_	_
7	Phosphodiesterase 4 D interacting protein	_	_	_	_	_	F1SB10	22	96,486	0.7	5.19		_	_	_	_
8	Adenosine deaminases, RNA specific B1	_	_	_	_	_	_	_	_	_	_	F1N0T4	113	79,070	4	9.08
9	Receptor for activated C kinase 1	_	_	_	_	_	_	_	_	_	_	P63243	100	35,511	13.9	7.6
10	Serine/threonine-protein kinase 38	_	_	_	_	_	_	_	_	_	_	F1MBL1	65	54,533	11.8	6.67

Accession number in the NCBInr database.

AN: accession number; MW: molecular weight; IP: isoelectric points; CP: coverage percentage; S: score.

Peptide score is shown as −10log(p), where p is the probability that the observed match is a random event. MASCOT protein scores (combined MS and MS/MS spectra) > 22 in rabbit, protein score >23 in pig, and >24 in bovine, were considered statistically significant (p < .05).

Table 3. Differences of soybean agglutinin specific binding functional regulatory proteins in EMs of different animal species.

		Rabbit					Swine					Bovine
No.	Protein name	AN	S	MW (kDa)	CP (%)	PI	AN	S	MW (kDa)	CP (%)	PI	AN	S	MW/kDa	CP (%)	PI
1	Solute carrier family 4 member 1	G1SLY0	275	102,346	9.8	5.13	F1RQY6	85	104,116	5.9	5.49	_	_	_	_	_
2	Anion exchange protein	G1SLY0	65	102,346	2.5	5.13	_	_	_	_	_	_	_	_	_	_
3	Sodium channel protein	G1U7U1	35	229,912	1.5	5.51	_	_	_	_	_	_	_	_	_	_
4	Cullin 9	G1SIL5	25	291,709	0.6	5.34	_	_	_	_	_	_	_	_	_	_
5	Rap guanine nucleotide exchange factor like 1	G1TJY0	25	71,524	1.9	5.91	_	_	_	_	_	_	_	_	_	_
6	Family with sequence similarity 71 member B	G1SHD8	24	59,557	3.1	9.64	_	_	_	_	_	_	_	_	_	_
7	Calmodulin regulated spectrin associated protein 1	_	_	_	_	_	F1S018	23	168,082	1	6.67	_	_	_	_	_
8	Heat shock 70 kDa protein 1 A	_	_	_	_	_	_	_	_	_	_	Q27975	169	70,500	16.1	5.68
9	T-complex protein 1 subunit beta	_	_	_	_	_	_	_	_	_	_	Q3ZBH0	97	57,781	2.8	6.18
10	78 kDa glucose-regulated protein	_	_	_	_	_	_	_	_	_	_	Q0VCX2	69	72,470	4.4	5.07

Accession number in the NCBInr database.

AN: accession number; MW: molecular weight; IP: isoelectric points; CP: coverage percentage; S: score.

Peptide score is shown as −10log(p), where p is the probability that the observed match is a random event. MASCOT protein scores (combined MS and MS/MS spectra) > 22 in rabbit, protein score >23 in pig, and >24 in bovine, were considered statistically significant (p < .05).

Figure 3. The overlapping and differential soybean agglutinin specific binding cytoskeleton proteins in three animal species.

Figure 4. The overlapping and differential soybean agglutinin specific binding catalytic enzyme proteins in three animal species.

Figure 5. The overlapping and differential soybean agglutinin functional regulatory proteins in three animal species.