Proteomic Difference Analysis of Whole Blood and Bloodstains

doi:10.12116/j.issn.1004-5619.2022.521204

Abstract

Abstract:

Objective To study the changes of protein levels in peripheral blood after it dried. Methods The proteins from whole blood and bloodstains were detected by liquid chromatography-tandem mass spectrometry （LC-MS/MS） and normalized by the label-free quantification （LFQ） method. The differential proteins were analyzed by using R 4.2.1 software, limma and edgeR package. The analysis of biological function, signaling pathway and subcellular localization for the differential proteins was then performed. Results A total of 623 and 596 proteins were detected in whole blood and bloodstains, respectively, of which 31 were statistically significant in the quantitative results, including 10 up-regulated and 21 down-regulated proteins in bloodstains. Conclusion The protein abundances in whole blood and bloodstains are highly correlated, and the variation of protein abundances may be related to the changes of endogenous and structural proteins in cells. The application of proteomics technology can assist the screening and identification of protein biomarkers, thereby introducing new biomarkers for forensic research.

Key words: forensic medicine, proteomics, liquid chromatography-tandem mass spectrometry （LC-MS/MS）, differential protein, label-free quantification method, whole blood, bloodstains

CLC Number:

Ao HUANG, Shu-bo WEN, Qian-qian KONG, Zhen-min ZHAO, Xi-ling LIU. Proteomic Difference Analysis of Whole Blood and Bloodstains[J]. Journal of Forensic Medicine, 2023, 39(6): 549-556.

Figures/Tables 5

References 44

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组别	样本	年龄/岁	平均年龄/岁	性别	血型	全血	血斑
P1	P1-1，P1-2，P1-3，P1-4，P1-5	25，25，25，26，28	26	女	O型	全血-1	血斑-1
P2	P2-1，P2-2，P2-3，P2-4，P2-5	25，26，27，29，47	30	男	O型	全血-2	血斑-2
P3	P3-1，P3-2，P3-3，P3-4，P3-5	25，26，26，26，28	26	男	A型	全血-3	血斑-3

组别	样本	年龄/岁	平均年龄/岁	性别	血型	全血	血斑
P1	P1-1，P1-2，P1-3，P1-4，P1-5	25，25，25，26，28	26	女	O型	全血-1	血斑-1
P2	P2-1，P2-2，P2-3，P2-4，P2-5	25，26，27，29，47	30	男	O型	全血-2	血斑-2
P3	P3-1，P3-2，P3-3，P3-4，P3-5	25，26，26，26，28	26	男	A型	全血-3	血斑-3

编号	蛋白质	Uniprot ID	基因	亚细胞定位	log₂FC	-log₁₀P	变化趋势
1	载脂蛋白C3	P02656	APOC3	分泌型	0.70	0.62	↑
2	核受体结合蛋白	Q9UHY1	NRBP1	胞质	0.72	1.52	↑
3	免疫球蛋白λ变量9-49	A0A0B4J1Y8	IGLV9-49	分泌型	26.00	10.15	↑
4	补体因子D	P00746	CFD	分泌型	23.57	11.08	↑
5	半胱氨酸蛋白酶抑制剂C	P01034	CST3	分泌型	23.62	10.01	↑
6	核糖核苷二磷酸还原酶大亚基	P23921	RRM1	胞质	22.95	10.90	↑
7	根蛋白	P35241	RDX	细胞膜	23.86	11.08	↑
8	肝细胞生长因子激活剂	Q04756	HGFAC	分泌型	22.96	9.24	↑
9	自噬相关蛋白3	Q9NT62	ATG3	胞质	23.03	11.08	↑
10	SEC14样蛋白4	Q9UDX3	SEC14L4	胞质	25.70	11.08	↑
11	铵转运体Rh A型	Q02094	RHAG	浆膜	-0.75	1.65	↓
12	胞质乙酰辅酶A硫脂水解酶	O00154	ACOT7	胞质	-23.63	10.64	↓
13	26S蛋白酶体非ATP酶调节亚基3	O43242	PSMD3	细胞质基质	-28.65	11.08	↓
14	α-辅肌动蛋白-4	O43707	ACTN4	细胞核	-24.56	8.25	↓
15	微管蛋白β链	P07437	TUBB	胞质	-23.18	10.93	↓
16	热激蛋白60	P10809	HSPD1	线粒体基质	-24.89	6.97	↓
17	断裂点丛集区蛋白	P11274	BCR	突触后致密区	-26.77	8.89	↓
18	延伸因子2	P13639	EEF2	胞质	-23.37	9.23	↓
19	连接桥粒斑珠蛋白	P14923	JUP	细胞连接处	-22.95	6.37	↓
20	异质性细胞核核糖蛋白A2/B1	P22626	HNRNPA2B1	细胞核	-23.07	8.36	↓
21	甘露聚糖结合凝集素丝氨酸蛋白酶1	P48740	MASP1	分泌型	-24.09	10.15	↓
22	X连锁Kx血型抗原	P51811	XK	内质网膜	-24.34	11.08	↓
23	Epiplakin	P58107	EPPK1	细胞骨架	-23.05	6.85	↓
24	转化蛋白RhoA	P61586	RHOA	细胞膜	-25.39	11.08	↓
25	免疫球蛋白结合蛋白1	P78318	IGBP1	胞质	-23.11	10.38	↓
26	1，4-α-葡聚糖分支酶	Q04446	GBE1	胞质	-22.60	11.08	↓
27	细胞间黏附分子4	Q14773	ICAM4	细胞膜	-24.15	11.08	↓
28	多聚C结合蛋白1	Q15365	PCBP1	细胞核	-24.62	10.22	↓
29	含NudC结构域的蛋白质2	Q8WVJ2	NUDCD2	染色体	-24.24	11.08	↓
30	羧肽酶B2	Q96IY4	CPB2	分泌型	-24.54	10.64	↓
31	Rabankyrin-5	Q9P2R3	ANKFY1	细胞膜	-22.81	11.08	↓

编号	蛋白质	Uniprot ID	基因	亚细胞定位	log₂FC	-log₁₀P	变化趋势
1	载脂蛋白C3	P02656	APOC3	分泌型	0.70	0.62	↑
2	核受体结合蛋白	Q9UHY1	NRBP1	胞质	0.72	1.52	↑
3	免疫球蛋白λ变量9-49	A0A0B4J1Y8	IGLV9-49	分泌型	26.00	10.15	↑
4	补体因子D	P00746	CFD	分泌型	23.57	11.08	↑
5	半胱氨酸蛋白酶抑制剂C	P01034	CST3	分泌型	23.62	10.01	↑
6	核糖核苷二磷酸还原酶大亚基	P23921	RRM1	胞质	22.95	10.90	↑
7	根蛋白	P35241	RDX	细胞膜	23.86	11.08	↑
8	肝细胞生长因子激活剂	Q04756	HGFAC	分泌型	22.96	9.24	↑
9	自噬相关蛋白3	Q9NT62	ATG3	胞质	23.03	11.08	↑
10	SEC14样蛋白4	Q9UDX3	SEC14L4	胞质	25.70	11.08	↑
11	铵转运体Rh A型	Q02094	RHAG	浆膜	-0.75	1.65	↓
12	胞质乙酰辅酶A硫脂水解酶	O00154	ACOT7	胞质	-23.63	10.64	↓
13	26S蛋白酶体非ATP酶调节亚基3	O43242	PSMD3	细胞质基质	-28.65	11.08	↓
14	α-辅肌动蛋白-4	O43707	ACTN4	细胞核	-24.56	8.25	↓
15	微管蛋白β链	P07437	TUBB	胞质	-23.18	10.93	↓
16	热激蛋白60	P10809	HSPD1	线粒体基质	-24.89	6.97	↓
17	断裂点丛集区蛋白	P11274	BCR	突触后致密区	-26.77	8.89	↓
18	延伸因子2	P13639	EEF2	胞质	-23.37	9.23	↓
19	连接桥粒斑珠蛋白	P14923	JUP	细胞连接处	-22.95	6.37	↓
20	异质性细胞核核糖蛋白A2/B1	P22626	HNRNPA2B1	细胞核	-23.07	8.36	↓
21	甘露聚糖结合凝集素丝氨酸蛋白酶1	P48740	MASP1	分泌型	-24.09	10.15	↓
22	X连锁Kx血型抗原	P51811	XK	内质网膜	-24.34	11.08	↓
23	Epiplakin	P58107	EPPK1	细胞骨架	-23.05	6.85	↓
24	转化蛋白RhoA	P61586	RHOA	细胞膜	-25.39	11.08	↓
25	免疫球蛋白结合蛋白1	P78318	IGBP1	胞质	-23.11	10.38	↓
26	1，4-α-葡聚糖分支酶	Q04446	GBE1	胞质	-22.60	11.08	↓
27	细胞间黏附分子4	Q14773	ICAM4	细胞膜	-24.15	11.08	↓
28	多聚C结合蛋白1	Q15365	PCBP1	细胞核	-24.62	10.22	↓
29	含NudC结构域的蛋白质2	Q8WVJ2	NUDCD2	染色体	-24.24	11.08	↓
30	羧肽酶B2	Q96IY4	CPB2	分泌型	-24.54	10.64	↓
31	Rabankyrin-5	Q9P2R3	ANKFY1	细胞膜	-22.81	11.08	↓

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