食品科学 ›› 2022, Vol. 43 ›› Issue (13): 1-7.doi: 10.7506/spkx1002-6630-20210327-339

• 基础研究 •    

促成骨细胞增殖活性骨胶原蛋白肽的靶向筛选及活性分析

陈永凯,郭玉杰,张鸿儒,刘泓,张春晖,姜珊   

  1. (1.中国农业科学院农产品加工研究所,农业农村部农产品加工综合性重点实验室,北京 100193;2.新疆泰昆集团有限责任公司,新疆 昌吉 831100;3.内蒙古阿荣旗牧原康肽实业有限公司,内蒙古 呼伦贝尔 162750)
  • 发布日期:2022-07-27
  • 基金资助:
    国家自然科学基金面上项目(32072156)

Targeted Screening and Activity Analysis of Bone Collagen Peptides with the Ability to Promote Osteoblast Proliferation

CHEN Yongkai, GUO Yujie, ZHANG Hongru, LIU Hong, ZHANG Chunhui, JIANG Shan   

  1. (1. Comprehensive Key Laboratory of Agro-products Processing, Ministry of Agriculture and Rural Affairs, Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 2. Xinjiang Taikun Group Co., Ltd., Changji 831100, China; 3. Inner Mongolia Arun Banner Muyuankangtai Industrial Co., Ltd., Hulun Buir 162750, China)
  • Published:2022-07-27

摘要: 酶法制备的骨胶原蛋白肽为混合肽,通常具有多种潜在的生物活性,但目前缺乏基于促成骨细胞增殖活性的靶向性研究。本实验以酶法制备的牦牛骨胶原蛋白肽为原料,采用高效液相色谱法与纳米液相色谱-串联质谱法分别测定混合肽的分子质量与序列构成,基于与表皮生长因子受体(epidermal growth factor receptor,EGFR)的CDOCKER半柔性对接,靶向筛选具有潜在促成骨细胞增殖活性的骨胶原蛋白肽,并进行体外细胞增殖培养验证。结果表明,共鉴定得到78 条牦牛骨胶原蛋白肽,主要以低分子质量肽段为主(小于3 000 Da的组分占91.8%),通过与EGFR对接发现,多肽GPAGPQGPRGDKGETGEQ(GP-18,1 736.815 Da)、TPEVDDEALEKFDK(TP-14,1 634.775 Da)、GPAGPQGPRGDKGETGE(GP-17,1 608.757 Da)、GKSGDRGETGPAGPAGPIGPV(GK-21,1 875.951 Da)和GKSGDRGETGPAGPAGPIGPVG(GK-22,1 932.973 Da)具有潜在促成骨细胞增殖活性。小鼠前成骨细胞系MC3T3-E1细胞培养实验结果表明,3.0 mg/mL GK-22促成骨细胞相对增殖率达106%,显著高于空白组(P<0.05)。本研究可为促成骨细胞增殖活性肽的靶向筛选和工业化制备提供理论参考。

关键词: 牦牛骨胶原蛋白肽;靶向筛选;成骨细胞;增殖活性

Abstract: Bone collagen peptides prepared by enzymatic hydrolysis are mixed peptides with a variety of potential biological activities. However, there is a lack of research on targeted screening of bone collagen peptides with the ability to promote osteoblast proliferation. In this study, the molecular mass and sequence of yak bone collagen peptides prepared by enzymatic hydrolysis were determined by high performance liquid chromatography (HPLC) and nano-liquid chromatography-tandem mass spectrometry (nano LC-MS/MS), respectively. The semi-flexible docking method CDOCKER based on epidermal growth factor receptor (EGFR) was used for the targeted screening of bone collagen peptides with the ability to promote osteoblast proliferation. Finally, in vitro cell tests were used to verify the molecular docking results. Results indicated that a total of 78 yak bone collagen peptides were identified, the low-molecular-mass ones being predominant (those with molecular mass less than 3 000 Da accounted for 91.8%). GPAGPQGPRGDKGETGEQ (GP-18, 1 736.815 Da), TPEVDDEALEKFDK (TP-14, 1 634.775 Da), GPAGPQGPRGDKGETGE (GP-17, 1 608.757 Da), GKSGDRGETGPAGPAGPIGPV (GK-21, 1 875.951 Da), and GKSGDRGETGPAGPAGPIGPVG (GK-22, 1 932.973 Da) had the potential to promote osteoblast proliferation. The relative proliferation rate of mouse MC3T3-E1 osteoblasts treated with 3.0 mg/mL GK-22 was 106%, which was significantly higher than that of the control group (P < 0.05). The results of this study provide a theoretical reference for targeted screening and industrial preparation of bioactive peptides with the ability to promote osteoblast proliferation.

Key words: yak bone collagen peptides; targeted screening; osteoblast; proliferation activity

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