Proteomic analysis of cyst fluid of Cysticercus cellulosae

doi:10.12140/j.issn.1000-7423.2019.02.005

Abstract

Abstract:

Objective The protein components of cyst fluid of Cysticercus cellulosae were analyzed by liquid chromatograph-mass spectrometer (LC-MS/MS) to identify the expression profile of C. cellulosae, The protein profile of Cysticercus fluid can be used to screen for the specific antigens as diagnostic markers and analyze the function of Cysticercus protein components involved in the invasion and survival of Cysticercus in the host. Methods The cystic fluid was aseptically collected from an infected pig and the protein components of the fluid was analyzed by LC-MS/MS after being digested with trypsin. The Blast2Go software was used for the comparison and annotation, and Gene Ontology (GO) was used to analyze classification of the annotated protein components. Results A total of 486 proteins were obtained by LC-MS/MS analysis, 158 of them were known proteins and the rest 328 were unknown proteins after being analyzed by searching with Mascot 2.2 software. The primary searching results identified that cystic fluid contains four highly abundant proteins (8 kDa glycoprotein, annexin, enolase, trypsin-like protein), and other specific antigens and a variety of enzymes, mainly located intracellularly and as cell components, of which annexin has binding functions mainly involved in a series of calcium-dependent membrane-type biological activities. The enolase has ionic binding function and hydrolase activity involved in the regulation of DNA-binding transcription factor and the glycolytic pathway. The results of GO analysis showed that 65 proteins were located intracellularly, 64 proteins located in the cell components, 25 proteins located in the organelles, and the rest located in the organelles, protein complexes, extracellular regions and cell membranes. In terms of molecular functions of identified proteins, 72 proteins have catalytic activities, 74 proteins have binding functions, and the remaining proteins are involved in transporter activities, molecular function regulators, structural molecular activities, etc. In terms of biological process classification, 69 proteins are involved in the cell process, 73 involved in the metabolic process, and 11 involved in the regulation of biological processes, signals, response to stimuli and cell localization. Conclusion The proteomic results of cyst fluid of Cysticercus identified a total of 486 known or unknown proteins, 4 of them are highly abundant proteins that can be used as candidate antigens for immunodiagnosis of cysticercosis. The identified enzymes may play important roles in the interaction between C. cellulosae and the host, providing a basis for further exploration of the invasion and parasitic mechanism of C. cellulosae.

Key words: Cysticercus cellulosae, Cyst fluid, LC-MS/MS, Diagnostic marker

CLC Number:

R383.34

Jian-da PANG, Yi-ning SONG, Xin-rui WANG, Ming-yuan LIU, Shu-min SUN. Proteomic analysis of cyst fluid of Cysticercus cellulosae[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2019, 37(2): 144-149.

Figures/Tables 4

Table 1

Fig. 1

Table 2

Fig. 2

References 28

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检索项目 Search item	参数Parameter
酶 Enzyme	Trypsin
数据库 Database	Uniprot Taenia
固定修饰 Fixed modifications	Carbamidomethyl
可变修饰 Variable modifications	Oxidation
肽质量耐受性 Peptide mass tolerance	0.002%
片段质量容限 Fragment mass tolerance	相对分子质量(M_r) 0.1
分数过滤 Filter by score	≥ 20

编号 No.	登录号 SwissProt/TrEMBL AC	蛋白名称 Protein name	肽段 Pep count	惟一肽段数 Unique pep count	肽段覆盖率/% Cover percent/%	相对分子质量 M_r	等电点 PI
1	Q9U8G9	囊尾蚴病10 kDa抗原 Cysticercosis 10 kDa antigen	88	8	43.53	9 650	8.89
2	Q8I0G1	排泄分泌抗原m13 Excretion-secretion antigen m13	52	8	50.59	9 820	9.75
3	E5LBB8	胰蛋白酶样蛋白质 Trypsin-like protein	33	17	39.96	5 412	7.09
4	D2U5C3	磷酸烯醇丙酮酸羧激酶Phosphoenolpyruvate carboxykinase	31	23	55.58	5 411	8.49
5	B6E4B3	8 kDa糖蛋白8 kDa glycoprotein	31	5	48.24	9 660	9.78
6	A0A1S5WII1	烯醇化酶Enolase	21	18	45.62	4 661	6.48
7	A0A0R3VSW8	果糖二磷酸醛缩酶 Fructose-bisphosphate aldolase	19	13	47.93	3 971	8.35
8	A0A291I2R3	丝氨酸蛋白酶抑制Serpin	17	15	41.90	4 554	7.63
9	A0A088FV33	胱抑素Cystatin	15	10	52.19	3 065	5.19
10	A0A0R3VUJ9	氨肽酶Aminopeptidase	13	12	11.57	11 410	6.88
11	K7SRY9	肌动蛋白Actin	11	9	30.59	4 174	5.30
12	A0A0R3VWJ0	磷酸甘油酸激酶 Phosphoglycerate kinase	10	7	17.59	4 413	7.56
13	A0A0R3W9G0	腺苷高半胱氨酸酶 Adenosylhomo cysteinase	5	5	12.21	5 182	6.16
14	Q967Z2	钙结合蛋白Calcium binding protein	5	5	17.47	4 557	4.41
15	A0A0R3WCA2	胺氧化酶Amine oxidase	5	4	4.90	8 099	6.55
16	Q5GM20	诊断抗原GP50GP50	5	4	23.51	2 957	8.68
17	Q8MPB9	小热休克蛋白 Small heat-shock protein	5	5	18.15	3 560	5.56
18	G8ZAG5	L-乳酸脱氢酶 L-lactate dehydrogenase	4	4	14.50	3 546	7.09
19	J9PE36	脂肪酸结合蛋白 Fatty acid binding protein	4	4	35.34	1 511	8.61
20	Q9XZL9	膜联蛋白Annexin	1	1	2.59	3 796	5.71