基于细胞膜色谱技术的鹿角胶肽段筛选及鉴定

doi:10.3976/j.issn.1002-4026.20230175

Abstract

Abstract:

Cell membrane chromatography/ultrahigh-phase liquid chromatography-mass spectrometry (CMC/UPLC-MS) was used to screen the active ingredients, i.e., peptides, in the traditional Chinese medicine tablet Cervi cornus Colla, and the obtained spectra were analyzed using Maxquant software, Perseus software, and Uniprot database. Structures of these peptides were identified using Protein Data Bank, and their molecular properties such as their biological activity, adverse reactions, relative molecular mass, isoelectric point, and stability index were predicted using a bioinformatics platform. With an activity probability of 0.09, the peptides were identified as nontoxic, nonhemolytic, sensitizing, and highly hydrophilic, with a relative molecular mass of 1 541.68, a peptide chain length of 14, an isoelectric point of 3.92, and an instability index of 34.39.This method provides a feasible research approach for rapidly screening and identifying active ingredients(e.g., peptides and proteins) that exhibit pharmacological effects.

Key words: Cervi Cornus Colla, cell membrane chromatography, bone marrow mesenchymal stem cells, interleukin-1β

CLC Number:

R284

SUN Tiefeng, ZHAO Yu, WANG Ping, DING Xianglong, DING Lijun, WANG Jinguo. To explore the pharmacodynamic substance basis of Cervi Cornus Colla based on cell membrane chromatography[J].Shandong Science, 2024, 37(5): 1-9.

Figures/Tables 10

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References 34

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药材/试剂	厂家	批号
DMEM-高糖液体培养基	美国赛默飞世尔公司	8123638
0.25%胰蛋白酶-EDTA溶液	NCM Biotech	20231130
青链霉素混合液	北京索莱宝科技有限公司	GP2308019
PBS缓冲液	Viva Cell	2352433
胎牛血清	ExCell Bio	12B234
鹿角片	山东建联盛嘉中药有限公司中药饮片厂	20210301
乙腈(色谱纯)	上海星可高纯溶剂有限公司	0113210806
乙酸(色谱纯)	天津市大茂化学试剂厂	20151221

仪器	厂家
96孔培养板	美国康宁公司
倒置生物显微镜	南京尼康江南光学仪器有限公司
CO₂培养箱	上海力申科学仪器有限公司
超净工作台	苏州安泰空气技术有限公司
DDL-5型离心机	上海安亭科技仪器厂
6 mL固相萃取柱	湖南比克曼生物科技有限公司
FW100型高速万能粉碎机	北京市永光明医疗仪器有限公司
十万分之一电子天平	德国塞多利斯科学仪器有限公司
高压液相色谱-质谱联用仪	美国赛默飞世尔科技公司
Water ACQUITY UPLC HSS C₁₈色谱柱(2.1 mm×100 mm,1.8 μm)	美国沃特世公司

数据库/软件	网址
Xcalibur软件(版本4.3)	https://thermo-scientific-xcalibur.software.informer.com/4.3/
Maxquant软件(版本2.4.0.0)	https://www.maxquant.org/
Perseus软件(版本2.0.10.0)	https://www.maxquant.org/perseus/
Peptide Ranker	http://distilldeep.ucd.ie/PeptideRanker/
ToxIBTL	https://server.wei-group.net/ToxIBTL/ProcessServlet
Hemopred	http://codes.bio/hemopred/
Algpred2.0	https://webs.iiitd.edu.in/raghava/algpred/submission.html
Uniprot	https://www.uniprot.org/
PBD	https://www1.rcsb.org/
Pepcalc.com-Peptide calculator	https://pepcalc.com-Peptide calculator
Expasy ProtParam tool	https://web.expasy.org/protparam

编号	序列	长度	相对分子质量
1	AGPSTGGRQGSRHEQAR	17	1 750.857 1
2	CCKAQEPEACFK	12	1 526.631 6
3	DSAQAATGECTATVAK	16	1 579.714 8
4	EIETYCLLIGGDDGACKSGGYKSK	24	2 620.214 9
5	GDTPTLQLEEVDPK	14	1 540.762 1
6	GTSVDGHSLASSNTDFALSLYK	22	2 269.086 3
7	GYEVHFFVNASDVSNVKAHLNASR	24	2 660.309 5
8	IECALNFAMQGAEEIR	16	1 850.865 5
9	NPSLLLGK	8	840.506 9
10	QLKKLHINYNNLTESVGPLPK	21	2 405.343 1
11	RHDFFKSAMPEGYVQER	17	2 095.989 8
12	SENFSSKTK	9	1 026.498 2
13	SEVAHRFKDLGEENFK	16	1 904.938 1
14	VSSVPSNSNVVVGTTNACAPSAR	23	2 273.107 0
15	VVNPTQA	7	727.386 5

序列	肽段	序列	肽段	序列	肽段	序列	肽段
10	MAPVPEPINE	80	MEKLRNSAYA	150	ALHLLSQEMS	220	RFVFYKTEIK
20	MMAYYSDENE	90	HVFHDDDLRN	160	REVVFCMSFV	230	DTVEFESVLY
30	LLFEADGPKQ	100	VLSFIFEEEP	170	QAEERDNKIP	240	PNWYISTSHP
40	MKSCIQHLDL	110	VIFETSSDEF	180	VALGIRDKNQ	250	EEKPVFLGHF
50	GSVEVGNIQL	120	LCDAAVQSVN	190	YLSCVKKGDT	260	RGGQDITDFR
60	QISHQLYNKS	130	CKLQDREQNS	200	PTLQLEEVDP		METLSP
70	FRQVVSVIVA	140	LVLASPCVLK	210	KVYPKRNMEK

To explore the pharmacodynamic substance basis of Cervi Cornus Colla based on cell membrane chromatography

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References 34

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序列	肽段	序列	肽段	序列	肽段	序列	肽段
10	PVQSIKCKLQ	50	FCMSFVQAEE	90	LEEVDPKVYP	130	ISTSQTEEKP
20	DREQKSLVLA	60	RDNKIPVALG	100	KRNMEKRFVF	140	VFLGHFRGGQ
30	SPCVLKALHL	70	IRDKNLYLSC	110	YKTEIKDTVE
40	LSQEMSREVV	80	VKKGDTPTLQ	120	FESVLYPNWY