Arginine kinase from Litopenaeus vannamei: Cloning, expression and catalytic properties

doi:10.1016/j.fsi.2009.02.012

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	Arginine kinase from Litopenaeus vannamei: Cloning, expression and catalytic properties
	Yao, Cui-Luan 2; Ji, Pei-Feng 2; Kong, Peng 2; Wang, Zhi-Yong 2; Xiang, Jian-Hai1
刊名	FISH & SHELLFISH IMMUNOLOGY
	2009-03-01
卷号	26 期号:3 页码:553-558
关键词	Arginine Kinase Cloning Expression Litopenaeus Vannamei Lps Activity
ISSN号	1050-4648
DOI	10.1016/j.fsi.2009.02.012
文献子类	Article
英文摘要	Arginine kinase (AK) is a phosphotransferase that plays a critical role in energy metabolism in invertebrates. in this paper, the full-length cDNA of AI( was cloned from shrimp, Litopenaeus vannamei by using RT-PCR and RACE PCR. It was 1446 bp encoding 356 amino acids, and belongs to the conserved phosphagen kinase family. The quantitative real-time reverse transcription PCR analysis revealed a broad expression of AK with the highest expression in the muscle and the lowest in the skin. The expression of AK after challenge with LIPS was tested in hemocytes and muscle, which indicated that the two peak values were 6.2 times (at 3 h) and 10.14 times (at 24 h) in the hemocytes compared with the control values, respectively (P < 0.05), while the highest expression of AK was 41 times (at 24 h) in the muscle compared with the control (P < 0.05). In addition, AK was expressed in Eschetichia coli by prokaryotic expression plasmid pGEX-4T-2. The recombinant protein was expressed as glutathione s-transferase (GST) arginine kinase (GST-AK) fusion protein, which was purified by affinity chromatography using Glutathione Sepharose 4B. After cleavage from GST by using a site-specific protease, the recombinant protein was identified by ESI-MS and showed AK activity. After treatment with 10 mM ATP, the enzyme activity significantly increased. However, the enzyme activity was inhibited by 10 mM alpha-ketoglutarate, 50 mM glucose and 200 mM ATP. This research suggested that AK might play an important role in the coupling of energy production and utilization and the immune response in shrimps. (C) 2009 Elsevier Ltd. All rights reserved.; Arginine kinase (AK) is a phosphotransferase that plays a critical role in energy metabolism in invertebrates. in this paper, the full-length cDNA of AI( was cloned from shrimp, Litopenaeus vannamei by using RT-PCR and RACE PCR. It was 1446 bp encoding 356 amino acids, and belongs to the conserved phosphagen kinase family. The quantitative real-time reverse transcription PCR analysis revealed a broad expression of AK with the highest expression in the muscle and the lowest in the skin. The expression of AK after challenge with LIPS was tested in hemocytes and muscle, which indicated that the two peak values were 6.2 times (at 3 h) and 10.14 times (at 24 h) in the hemocytes compared with the control values, respectively (P < 0.05), while the highest expression of AK was 41 times (at 24 h) in the muscle compared with the control (P < 0.05). In addition, AK was expressed in Eschetichia coli by prokaryotic expression plasmid pGEX-4T-2. The recombinant protein was expressed as glutathione s-transferase (GST) arginine kinase (GST-AK) fusion protein, which was purified by affinity chromatography using Glutathione Sepharose 4B. After cleavage from GST by using a site-specific protease, the recombinant protein was identified by ESI-MS and showed AK activity. After treatment with 10 mM ATP, the enzyme activity significantly increased. However, the enzyme activity was inhibited by 10 mM alpha-ketoglutarate, 50 mM glucose and 200 mM ATP. This research suggested that AK might play an important role in the coupling of energy production and utilization and the immune response in shrimps. (C) 2009 Elsevier Ltd. All rights reserved.
学科主题	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
URL标识	查看原文
语种	英语
WOS记录号	WOS:000265518300027
公开日期	2010-12-22
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/3057]
专题	海洋研究所_实验海洋生物学重点实验室
作者单位	1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 2.Jimei Univ, Coll Fisheries, Fisheries Biotechnol Inst, Key Lab Sci & Technol Aquaculture & Food Safety F, Xiamen 361021, Peoples R China
推荐引用方式 GB/T 7714	Yao, Cui-Luan,Ji, Pei-Feng,Kong, Peng,et al. Arginine kinase from Litopenaeus vannamei: Cloning, expression and catalytic properties[J]. FISH & SHELLFISH IMMUNOLOGY,2009,26(3):553-558.
APA	Yao, Cui-Luan,Ji, Pei-Feng,Kong, Peng,Wang, Zhi-Yong,&Xiang, Jian-Hai.(2009).Arginine kinase from Litopenaeus vannamei: Cloning, expression and catalytic properties.FISH & SHELLFISH IMMUNOLOGY,26(3),553-558.
MLA	Yao, Cui-Luan,et al."Arginine kinase from Litopenaeus vannamei: Cloning, expression and catalytic properties".FISH & SHELLFISH IMMUNOLOGY 26.3(2009):553-558.