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学者姓名：潘剑茹
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Abstract ：
目的 探讨前导肽的融合对人锰超氧化物歧化酶(SOD2)结构以及抗顺铂(DDP)诱导的肾损伤效应.方法 通过结构预测和SOD比活力测定分析线粒体靶向序列(MTS)对SOD2构效的影响;建立昆明(KM)小鼠DDP损伤模型,以阿米福汀(AMFT)为阳性对照,测定小鼠肾功能、肾脏指数、肾脏抗氧化能力,同时观察肾脏表观及病理变化,以评估MTS-SOD2抗DDP诱导的肾损伤效应.结果 MTS前导肽对SOD2的二三级结构有一定影响,但也使MTS-SOD2蛋白的比活力提高.预给予中剂量MTS-SOD2(0.84 mg/kg)可以使损伤鼠肾脏丙二醛(MDA)水平显著降低,SOD活力和总抗氧化能力(T-AOC)显著增加,进而减少肾脏病理损伤,维持肾功能,整体效果与200 mg/kg AMFT相当甚至优于后者.结论 MTS前导肽既增强了SOD2的活性,又使其因具有跨膜功能而发挥优异的抗DDP诱导的肾损伤效应.
Keyword ：
前导肽 前导肽 肾损伤 肾损伤 锰超氧化物歧化酶 锰超氧化物歧化酶 顺铂 顺铂
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| GB/T 7714 | 潘剑茹 , 韩亚南 , 何夏琪 et al. 带前导肽的人锰超氧化物歧化酶抗顺铂诱导的肾损伤效应 [J]. | 肿瘤防治研究 , 2023 , 50 (7) : 675-680 . |
| MLA | 潘剑茹 et al. "带前导肽的人锰超氧化物歧化酶抗顺铂诱导的肾损伤效应" . | 肿瘤防治研究 50 . 7 (2023) : 675-680 . |
| APA | 潘剑茹 , 韩亚南 , 何夏琪 , 叶小强 , 何火聪 . 带前导肽的人锰超氧化物歧化酶抗顺铂诱导的肾损伤效应 . | 肿瘤防治研究 , 2023 , 50 (7) , 675-680 . |
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Annexin A2 (ANXA2) has been found to be involved in cancer proliferation, metastasis and prognosis; however, its exact role in nasopharyngeal carcinoma (NPC) radioresistance remains unknown. We found that ANXA2 expression was correlated with prognosis in NPC patients, and longer overall survival in NPC patients with low ANXA2 expression than those with high ANXA2 expression. ANXA2 knockdown increased the radiosensitivity in radioresistant NPC cells, and ANXA2 overexpression decreased the radiosensitivity in NPC cells. Knocking-down ANXA2 expression increased the irradiation-induced apoptosis of radioresistant NPC cells, and ANXA2 overexpression decreased the irradiation-induced apoptosis of NPC cells. ANXA2 knockdown induced G2/M phase arrest in NPC cells post-irradiation, and ANXA2 overexpression abrogated G2/M phase arrest in NPC cells post-irradiation. ANXA2 overexpression resulted in inhibition of the p38 MAPK-HSP27 pathway, while ANXA2 knockdown resulted in activation of the p38 MAPK-HSP27 pathway. In addition, ANXA2 knockdown increased the radiosensitivity of the xenografted tumors in nude mice. Our data demonstrate that knockdown of Annexin A2 enhanced radiosensitivity in NPC by increasing G2/M-phase arrest, apoptosis and activating the p38 MAPK-HSP27 pathway. ANXA2 may be a promising target used to overcome radioresistance in NPC.
Keyword ：
annexin A2 annexin A2 apoptosis apoptosis cell-cycle arrest cell-cycle arrest nasopharyngeal carcinoma nasopharyngeal carcinoma radioresistance radioresistance
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| GB/T 7714 | He, Huocong , Lin, Keyu , Zou, Changyan et al. Knockdown of Annexin A2 Enhances Radiosensitivity by Increasing G2/M-Phase Arrest, Apoptosis and Activating the p38 MAPK-HSP27 Pathway in Nasopharyngeal Carcinoma [J]. | FRONTIERS IN ONCOLOGY , 2022 , 12 . |
| MLA | He, Huocong et al. "Knockdown of Annexin A2 Enhances Radiosensitivity by Increasing G2/M-Phase Arrest, Apoptosis and Activating the p38 MAPK-HSP27 Pathway in Nasopharyngeal Carcinoma" . | FRONTIERS IN ONCOLOGY 12 (2022) . |
| APA | He, Huocong , Lin, Keyu , Zou, Changyan , Pan, Jianru , Fu, Wankai , Zhou, Yan et al. Knockdown of Annexin A2 Enhances Radiosensitivity by Increasing G2/M-Phase Arrest, Apoptosis and Activating the p38 MAPK-HSP27 Pathway in Nasopharyngeal Carcinoma . | FRONTIERS IN ONCOLOGY , 2022 , 12 . |
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Antioxidant enzymes fused with cell-penetrating peptides could enter cells and protect cells from irradiation damage. However, the unselective transmembrane ability of cell-penetrating peptide may also bring antioxidant enzymes into tumor cells, thus protecting tumor cells and consequently reducing the efficacy of radiotherapy. There are active matrix metalloproteinase (MMP)-2 or MMP-9 in most tumor cellular microenvironments. Therefore, a fusion protein containing an MMP-2/9 cleavable substrate peptide X, a cell-penetrating peptide R9, a glutathione S-transferase (GST), and a human Cu, Zn superoxide dismutase (SOD1), was designed and named GST-SOD1-X-R9. In the tumor microenvironment, GST-SOD1-X-R9 would lose its cell-penetrating peptide and could not enter tumor cells due to the cleavage of substrate X by active MMP-2/9, thereby achieving selected entering normal cells. The complete nucleotide sequence of SOD1-X-R9 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1. The pGEX4T-1-SOD1-X-R9 recombinant plasmid was obtained, and soluble expression of the fusion protein was achieved. GST-SOD1-X-R9 was purified by ammonium sulfate precipitation and GST affinity chromatography. The molecular weight of the fusion protein was approximately 47 kDa, consistent with the theoretical value. The SOD and GST activities were 2 954 U/mg and 328 U/mg, respectively. Stability test suggested that almost no change in either SOD activity or GST activity of GST-SOD1-X-R9 was observed under physiological conditions. The fusion protein could be partially digested by collagenase IV in solution. Subsequently, the effect of MMP-2/9 activity on transmembrane ability of the fusion protein was tested using 2D and 3D cultured HepG2 cells. Little extracellular MMP-2 activity of HepG2 cells was observed under 2D culture condition. While under the 3D culture model, the size and the MMP-2 activity of the HepG2 tumor spheroid increased daily. GST-SOD1-R9 proteins showed the same transmembrane efficiency in 2D cultured HepG2 cells, but the transmembrane efficiency of GST-SOD1-X-R9 in 3D cultured HepG2 spheres was reduced remarkably. This study provided a basis for further investigating the selectively protective effect of GST-SOD1-X-R9 against oxidative damage in normal cells. ©2022 Chin J Biotech, All rights reserved.
Keyword ：
antioxidant enzyme antioxidant enzyme cell permeable cell permeable expression in Escherichia coli expression in Escherichia coli MMP-2/9 sensitive MMP-2/9 sensitive purification purification stability stability
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| GB/T 7714 | He, H. , Lin, L. , Li, L. et al. Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征] [J]. | Chinese Journal of Biotechnology , 2022 , 38 (9) : 3515-3527 . |
| MLA | He, H. et al. "Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征]" . | Chinese Journal of Biotechnology 38 . 9 (2022) : 3515-3527 . |
| APA | He, H. , Lin, L. , Li, L. , Wu, L. , Lin, H. , Pan, J. . Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征] . | Chinese Journal of Biotechnology , 2022 , 38 (9) , 3515-3527 . |
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融合了跨膜肽的抗氧化酶可进入细胞,保护细胞免受放射损伤.然而跨膜肽的跨膜能力没有靶向性,其也可把抗氧化酶带入肿瘤细胞进而保护肿瘤细胞,降低放疗的效果.为此,根据多数肿瘤细胞微环境中存在活性基质金属蛋白酶(matrix metalloproteinase,MMP)-2或MMP-9的特点,在细胞跨膜肽R9与人铜、锌超氧化物歧化酶(superoxide dismutase 1,SOD1)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)之间融合 MMP-2/9的底物肽X,设计了融合蛋白GST-SOD1-X-R9.该蛋白在肿瘤微环境中可因MMP-2/9酶切底物肽X而失去跨膜肽,从而无法进入肿瘤细胞,进而只能进入正常细胞.全基因合成SOD1-X-R9序列,并将其插入原核表达载体pGEX-4T-1中,得到表达质粒,并实现了GST-SOD1-X-R9融合蛋白的可溶表达.GST-SOD1-X-R9经硫酸铵沉淀和GST亲和层析纯化,分子量约为47kDa,与理论值一致.纯化的融合蛋白的SOD活性和GST活性分别为2 954 U/mg和328 U/mgoGST-SOD1-X-R9的SOD活性或GST活性在生理条件下几乎没有变化.该融合蛋白在溶液中可被胶原酶Ⅳ部分水解.分别建立了2D和3D培养的HepG2细胞模型来检验肿瘤微环境中的MMP-2活力对该蛋白跨膜能力的影响.在2D培养模型中,HepG2的MMP-2活力极低,但在3D培养模型中,随着培养时间的增加,HepG2肿瘤球的体积变大,其胞外MMP-2活力也随之增强.GST-SOD1-X-R9在2D培养的HepG2细胞中具有和GST-SOD1-R9蛋白一样的跨膜效率,但在3D培养的HepG2细胞球中的跨膜能力大大降低.本研究为后续深入研究GST-SOD1-X-R9靶向防护正常细胞的氧化损伤效应奠定了基础.
Keyword ：
MMP-2/9敏感性 MMP-2/9敏感性 可跨膜 可跨膜 大肠杆菌表达体系 大肠杆菌表达体系 抗氧化酶 抗氧化酶 稳定性 稳定性 纯化 纯化
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| GB/T 7714 | 何火聪 , 林丽香 , 李玲玲 et al. 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征 [J]. | 生物工程学报 , 2022 , 38 (9) : 3515-3527 . |
| MLA | 何火聪 et al. "基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征" . | 生物工程学报 38 . 9 (2022) : 3515-3527 . |
| APA | 何火聪 , 林丽香 , 李玲玲 , 吴伦巧 , 林海英 , 潘剑茹 . 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征 . | 生物工程学报 , 2022 , 38 (9) , 3515-3527 . |
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Abstract ：
融合了跨膜肽的抗氧化酶可进入细胞，保护细胞免受放射损伤。然而跨膜肽的跨膜能力没有靶向性，其也可把抗氧化酶带入肿瘤细胞进而保护肿瘤细胞，降低放疗的效果。为此，根据多数肿瘤细胞微环境中存在活性基质金属蛋白酶(matrix metalloproteinase,MMP)-2或MMP-9的特点，在细胞跨膜肽R9与人铜、锌超氧化物歧化酶(superoxide dismutase 1,SOD1)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)之间融合MMP-2/9的底物肽X，设计了融合蛋白GST-SOD1-X-R9。该蛋白在肿瘤微环境中可因MMP-2/9酶切底物肽X而失去跨...
Keyword ：
MMP-2/9敏感性 MMP-2/9敏感性 可跨膜 可跨膜 大肠杆菌表达体系 大肠杆菌表达体系 抗氧化酶 抗氧化酶 稳定性 稳定性 纯化 纯化
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| GB/T 7714 | 何火聪 , 林丽香 , 李玲玲 et al. 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征（英文） [J]. | 生物工程学报 , 2022 , 38 (09) : 3515-3527 . |
| MLA | 何火聪 et al. "基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征（英文）" . | 生物工程学报 38 . 09 (2022) : 3515-3527 . |
| APA | 何火聪 , 林丽香 , 李玲玲 , 吴伦巧 , 林海英 , 潘剑茹 . 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征（英文） . | 生物工程学报 , 2022 , 38 (09) , 3515-3527 . |
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Background: Paraspeckle component 1 (PSPC1) is overexpressed in various cancer and correlated with poor survival in the patients. However, little is known about its expression and role in the progression of nasopharyngeal carcinomas (NPC). The purpose of this study is to examine PSPC1 expression in NPC and explore its role in clinical prognosis of radiation therapy. Methods: The association of PSPC1 expression with clinicopathological features of 109 NPC patients was examined using partial correlation analysis. Cancer tissues were obtained prior to clinical treatment. All cases were diagnosed and pathologically confirmed to be poorly differentiated or undifferentiated NPC without distant metastasis. The patients were then treated with radiation and followed-up. Survival analysis was performed. Results: Partial correlation analysis revealed that the PSPC1 expression in NPC was correlated with N classification, recurrence, prognosis and radiosensitivity in NPC patients, but not with the gender, age, pathohistological pattern, clinical stage, and T classification. The overexpression of PSPC1 was detected in 64 samples (58.72%). Kaplan-Meier survival analysis revealed that the overall survival (OS) was longer in NPC patients with PSPC1 low expression than that in those with PSPC1 high expression. Moreover, patients with the overexpression of PSPC1 had a low progression-free survival and distant metastasis-free survival rate, compared to those who had a low expression of PSPC1. Although not statistically significant, patients with high expression of PSPC1 had a lower locoregional recurrence-free survival rate than those with low expression, and the curves between the two groups was well separated. Conclusion: PSPC1 overexpression was associated with poor prognosis for NPC, which might be a novel useful biomarker to predict the response of NPC to radiation therapy and its clinical outcome.
Keyword ：
clinical prognosis clinical prognosis nasopharyngeal carcinoma nasopharyngeal carcinoma overexpression overexpression paraspeckle component 1 paraspeckle component 1 PSPC1 PSPC1 radiation radiation
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| GB/T 7714 | He, Huocong , Zhang, Lurong , Lin, Keyu et al. The Prognosis Value of PSPC1 Expression in Nasopharyngeal Cancer [J]. | CANCER MANAGEMENT AND RESEARCH , 2021 , 13 : 3281-3291 . |
| MLA | He, Huocong et al. "The Prognosis Value of PSPC1 Expression in Nasopharyngeal Cancer" . | CANCER MANAGEMENT AND RESEARCH 13 (2021) : 3281-3291 . |
| APA | He, Huocong , Zhang, Lurong , Lin, Keyu , Huang, Zhengrong , Zhou, Yan , Lin, Shaojun et al. The Prognosis Value of PSPC1 Expression in Nasopharyngeal Cancer . | CANCER MANAGEMENT AND RESEARCH , 2021 , 13 , 3281-3291 . |
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为了进一步研究当归(Angelica sinensis)生药中的蛋白质及其功能,通过80％硫酸铵沉淀、Sephadex G-50凝胶过滤层析、DEAE-Sepharose阴离子交换层析,首次从当归生药中纯化出两种分子量相近的蛋白(命名为ASPR-C-1和ASPR-C-2).ASPR-C-1和ASPR-C-2在SDS-PAGE上的分子量分别为17.33 kDa和17.18 kDa,在溶液中主要以单体形式存在,但会部分形成二聚体,二者均为糖蛋白,糖基含量分别为2.6％和8.2％.经基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-TOFTM)鉴定发现ASPR-C-1和ASPR-C-2均为病程相关(Pathogenesis-related 10,PR-10)家族蛋白,且具有核糖核酸酶活性,比活分别为73.60 U/mg和146.76 U/mg.两种蛋白的最适pH相近,均为5.6左右,但最适温度不同,ASPR-C-1的为50℃,ASPR-C-2的为60℃.二者虽然在60℃下都表现出最大的酶活力稳定性,但在更高的处理温度(80-100℃)下,ASPR-C-1迅速失活,最终仅余20％左右活力,ASPR-C-2则表现出良好的热稳定性,最终仍有80％左右活力.此外,Fe2+对二者的酶活性具有激活作用,而Ca2+、Mg2+、Zn2+、Mn2+、Ag+、Cu2+、EDTA、DTT和SDS则会不同程度地抑制二者的酶活性.研究结果为深入研究来自当归生药的PR-10蛋白的生物学功能奠定了基础.
Keyword ：
PR-10 PR-10 当归生药 当归生药 核糖核酸酶 核糖核酸酶 糖蛋白 糖蛋白 纯化 纯化
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| GB/T 7714 | 王香玲 , 李娴 , 何火聪 et al. 当归生药中两种PR-10蛋白亚型的纯化与表征 [J]. | 生物工程学报 , 2019 , 35 (1) : 159-168 . |
| MLA | 王香玲 et al. "当归生药中两种PR-10蛋白亚型的纯化与表征" . | 生物工程学报 35 . 1 (2019) : 159-168 . |
| APA | 王香玲 , 李娴 , 何火聪 , 李玲玲 , 吕迪 , 陈翠煌 et al. 当归生药中两种PR-10蛋白亚型的纯化与表征 . | 生物工程学报 , 2019 , 35 (1) , 159-168 . |
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目的探讨葡萄糖调节蛋白GRP78蛋白表达与鼻咽癌临床特征及预后的关系。方法随机选取2012年2月至2012年7月福建省肿瘤医院初诊鼻咽癌患者的治疗前鼻咽肿瘤活检组织标本107例,中位年龄47岁,全部病例鼻咽部组织均经病理学确诊且无远处转移。免疫组织化学法检测GRP78蛋白在鼻咽癌肿瘤组织的表达水平。根据鼻咽癌患者的性别、年龄、病理类型、肿瘤分期、无远处转移生存时间和总生存时间等临床资料,分析GRP78蛋白表达与临床分期、肿瘤远处转移和总生存等的关系。结果全部鼻咽肿瘤活检组织标本中GRP78蛋白的阳性表达率为72.9%(78/107)。GRP78蛋白的表达水平与患者的性别、年龄、病理类型、临床分...
Keyword ：
GRP78 GRP78 临床分期 临床分期 临床意义 临床意义 低表达 低表达 活检组织 活检组织 高表达 高表达 鼻咽癌患者 鼻咽癌患者 鼻咽肿瘤 鼻咽肿瘤
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| GB/T 7714 | 何火聪 , 苏颖 , 林可焴 et al. GRP78在鼻咽癌组织的表达及其临床意义 [C] //2018年中国肿瘤标志物学术大会暨第十二届肿瘤标志物青年科学家论坛论文集 . 2018 . |
| MLA | 何火聪 et al. "GRP78在鼻咽癌组织的表达及其临床意义" 2018年中国肿瘤标志物学术大会暨第十二届肿瘤标志物青年科学家论坛论文集 . (2018) . |
| APA | 何火聪 , 苏颖 , 林可焴 , 陈超 , 邹长棪 , 潘剑茹 . GRP78在鼻咽癌组织的表达及其临床意义 2018年中国肿瘤标志物学术大会暨第十二届肿瘤标志物青年科学家论坛论文集 . (2018) . |
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Background/Aims: Nasopharyngeal carcinoma (NPC) is rare worldwide but remains highly prevalent in endemic regions, notably in southern China. Radiotherapy remains the treatment of choice for NPC, but radioresistance has been identified as a major cause of therapeutic failure. The Wnt/-catenin signaling has been found to be involved in NPC radioresistance; however, the effect of -catenin overexpression on radioresistance remains unknown in NPC until now. This study aimed to examine the impact of -catenin overexpression on the radiosensitivity of human NPC CNE-2 cells. Methods: Immunohistochemistry was performed to detect the -catenin expression in normal nasopharyngeal specimens and NPC specimens. The human NPC CNE-2 cell line overexpressing -catenin was modeled by transfection with the pcDNA3.1/Hygro(+)/-catenin recombinant vector (transfection group), while cells transfected with the pcDNA3.1/Hygro(+) vector served as negative controls and non-transfected cells served as blank controls. The expression of key molecules of the Wnt/-catenin signaling pathway was determined using Western blotting and qPCR assays, and the changes of radiation sensitivity were measured with a colony-formation assay. Cell viability was measured by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 -diphenyltetrazolium bromide) assay. In addition, the cell cycle and apoptosis was detected using flow cytometry and the TCF/LEF transcriptional activity was measured with a Dual Luciferase Reporter Assay System. Results: Immunohistochemical staining showed high -catenin expression in radioresistant NPC specimens, and low expression in radiosensitive NPC specimens and normal nasopharyngeal specimens. Western blotting and qPCR assays detected higher -catenin expression in the transfection group than in the negative and blank controls (P < 0.01). Down-regulation of GSK-3 expression (P < 0.05) and up-regulation of Cyclin D1 expression (P < 0.01) was detected in -catenin overexpressing NPC cells exposed to X-ray radiation relative to negative and blank controls. Colony-formation assay revealed higher D-0, D-q and SF in the transfection group than in the negative and blank control groups post-radiation, and the SER in the transfection group was 0.75-fold and 0.68-fold greater than that in the blank and negative control groups, respectively. MTT assay revealed that the viability of CNE-2 cells was significantly higher in the transfection group (96% +/- 8.72%) than in the negative control group (74.67 +/- 7.05%) and the blank control group (75.33% +/- 7.02%) 24 h post-exposure to 6 Gy X-ray radiation (P < 0.05). X-ray radiation led to a lower proportion of CNE-2 cells at the G2/M phase and a lower apoptotic rate in the transfection group than in the negative and blank control groups (P < 0.05). In addition, the TCF/LEF transcriptional activity was higher in the transfection group than in the negative and blank control groups (P < 0.01), and 6 Gy X-ray radiation elevated the TCF/LEF transcriptional activity relative to 0 Gy radiation in the transfection group (P < 0.01). Conclusion: -catenin overexpression may decrease the radiation sensitivity in NPC CNE-2 cells through activating the downstream transcriptional factors of -catenin, and reducing G2/M arrest and cell apoptosis.
Keyword ：
-catenin -catenin Nasopharyngeal carcinoma Nasopharyngeal carcinoma Overexpression Overexpression Radiosensitivity Radiosensitivity
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| GB/T 7714 | He, Huocong , Lin, Keyu , Su, Ying et al. Overexpression of -Catenin Decreases the Radiosensitivity of Human Nasopharyngeal Carcinoma CNE-2 Cells [J]. | CELLULAR PHYSIOLOGY AND BIOCHEMISTRY , 2018 , 50 (5) : 1929-1944 . |
| MLA | He, Huocong et al. "Overexpression of -Catenin Decreases the Radiosensitivity of Human Nasopharyngeal Carcinoma CNE-2 Cells" . | CELLULAR PHYSIOLOGY AND BIOCHEMISTRY 50 . 5 (2018) : 1929-1944 . |
| APA | He, Huocong , Lin, Keyu , Su, Ying , Lin, Shaojun , Zou, Changyan , Pan, Jianru et al. Overexpression of -Catenin Decreases the Radiosensitivity of Human Nasopharyngeal Carcinoma CNE-2 Cells . | CELLULAR PHYSIOLOGY AND BIOCHEMISTRY , 2018 , 50 (5) , 1929-1944 . |
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In this study, two pathogenesis-related (PR) class 10 protein isoforms, ASPR-1 and ASPR-2, were purified from fresh roots of the Chinese medicinal plant Angelica sinensis (A. sinensis) using 80% ammonium sulfate precipitation, Sephadex G50 gel filtration chromatography, and DEAE-Sepharose ion-exchange chromatography. The molecular masses of ASPR-1 and ASPR-2 were estimated to be 16.66 kDa and 16.46 kDa, respectively, using sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The isoforms are both glycoproteins containing glycosyl contents of 1.8% (ASPR-1) and 3.4% (ASPR-2). The two isoforms were predominantly present as monomers, but they partially dimerized in solution. The 15 N-terminal amino acids of ASPR-1 were determined to be GIQKTEVEAPSTVSA, with significant sequence homology to certain PR-10 proteins. ASPR-2 was also identified using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) analysis to be a PR-10 protein. The isoforms both exhibited ribonuclease (RNase) activity, with ASPR-2 having higher specific activity (128.85 U mg(-1)) than ASPR-1 (68.67 U mg(-1)). The isoforms had the same optimal temperature of 50 degrees C but different optimal pH values of 5.0 (ASPR-1) and 6.0 (ASPR-2). The RNase activities of the isoforms were both stable for 30 min at 50 degrees C, rapidly decreasing at higher or lower processing temperatures. However, ASPR-1 retained higher residual activity (89.4%-80.9%) than ASPR-2 (74.3%-67.9%) at temperatures from 40 degrees C to 60 degrees C. These results provide additional information to enrich the current knowledge of poorly annotated A. sinensis proteins.
Keyword ：
Angelica sinensis Angelica sinensis Glycoprotein Glycoprotein Isoform Isoform Pathogenesis-related (PR) class 10 protein Pathogenesis-related (PR) class 10 protein Purification Purification Ribonuclease activity Ribonuclease activity
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| GB/T 7714 | Pan, Jianru , Wang, Xiangling , Li, Lingling et al. Purification and characterization of two pathogenesis-related class 10 protein isoforms with ribonuclease activity from the fresh Angelica sinensis roots [J]. | PLANT PHYSIOLOGY AND BIOCHEMISTRY , 2018 , 128 : 66-71 . |
| MLA | Pan, Jianru et al. "Purification and characterization of two pathogenesis-related class 10 protein isoforms with ribonuclease activity from the fresh Angelica sinensis roots" . | PLANT PHYSIOLOGY AND BIOCHEMISTRY 128 (2018) : 66-71 . |
| APA | Pan, Jianru , Wang, Xiangling , Li, Lingling , Li, Xian , Ye, Xiaoqiang , Lv, Di et al. Purification and characterization of two pathogenesis-related class 10 protein isoforms with ribonuclease activity from the fresh Angelica sinensis roots . | PLANT PHYSIOLOGY AND BIOCHEMISTRY , 2018 , 128 , 66-71 . |
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