来自血,Tegillarca granosa 的多子单元血红蛋白的可溶性表达和功能特征
Weifeng Zhang1, Tiancheng Chen2, Yazhuo Hu3
1School of Marine Science, Ningbo University, Ningbo 315832, China; Key Laboratory of Aquatic Germplasm Resource of Zhejiang, College of Biological & Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China; Ninghai Institute of Mariculture Breeding and Seed Industry, Zhejiang Wanli University, Ningbo 315604, China.
概括
生产无脊椎动物复合血红蛋白 (rHb) 是一个挑战. 这项研究成功地表达和净化了T. granosa rHbs,显示了可比的氧结合和增强的酸盐还原酶活性,为无脊椎动物Hb研究提供了新的框架.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 结构生物学 结构生物学
背景情况:
- 研究血红蛋白 (Hb) 的结构功能关系至关重要.
- 生产生物活跃的无脊椎动物重组Hb (rHb),特别是多重体形式,存在重大挑战.
- 大肠杆菌表达系统为重组蛋白质生产提供了一个强大的工具.
研究的目的:
- 描述来自血Tegillarca granosa的同位体rHbI和异位体rHbII的重组表达,净化和特征.
- 优化表达条件,以获得高产量的可溶性T. granosa rHbs.
- 为了比较复合和原生T. granosa Hbs. 的功能性质.
主要方法:
- 在Escherichia coli (JM109(DE3) 菌株) 中的重组表达与优化诱导条件 (15°C,0.25 mM IPTG) 和子优化.
- 使用已确定的染色学技术净化rHbs.
- 使用循环二重化 (CD) 光谱,紫外可见 (UV-Vis) 光谱,氧结合试验和化物减少试验进行表征.
主要成果:
- 在优化条件下,可溶性T. granosa rHbs的高产量 (65.4-135.6 g/L) 得到了实现.
- 纯化的rHbs表现出类似于原生Hbs (nHbs) 的二次结构和联结能力.
- rHbI和rHbII与nHbs具有相似的氧系亲和力,但酸还原酶活性明显高于nHbs.
结论:
- 已建立的复合表达和净化方法为研究T. granosa Hb和其他无脊椎动物Hbs.提供了有价值的框架.
- 重组表达克服了生产功能性无脊椎动物多重体Hbs.Hbs.的挑战.
- 在rHbs中增强的酸还原酶活性表明潜在的结构性或N端标签影响.
相关概念视频
Gene Families
9.2K
Gene families consist of groups of genes proposed to have originated from a common ancestor. Typically these arise through events in which a gene or genes are mistakenly duplicated during cell division. Unlike their parent genes (which are subject to selection pressure to maintain function), these gene copies do not need to preserve their sequences and may evolve at a relatively faster rate.
Occasionally these regions can be adapted to take on new roles within the organism, becoming novel genes...
Occasionally these regions can be adapted to take on new roles within the organism, becoming novel genes...
9.2K
Globular and Fibrous Proteins
44.5K
Many proteins can be classified into two distinct subtypes - globular or fibrous. These two types differ in their shapes and solubilities.
Globular proteins are also known as spheroproteins and typically are approximately round in shape. They contain a mix of amino acid types and contain differing sequences in their primary structures. Globular proteins have many different functions, such as enzymes, cellular messengers, and molecular transporters. These roles often require the proteins to be...
Globular proteins are also known as spheroproteins and typically are approximately round in shape. They contain a mix of amino acid types and contain differing sequences in their primary structures. Globular proteins have many different functions, such as enzymes, cellular messengers, and molecular transporters. These roles often require the proteins to be...
44.5K
General Transcription Factors
5.6K
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
5.6K
Hemoglobin
4.5K
Hemoglobin is a globular protein made up of four subunits. Two of these subunits are alpha chains, and the other two are beta chains. Each subunit contains a molecule of heme, which has an iron atom and can bind to oxygen. When an oxygen molecule binds to one heme group, it changes the shape of hemoglobin, making it easier for the other heme groups to bind oxygen as well.
When all four heme groups are bound to oxygen, the resulting molecule is called oxyhemoglobin. As a result, arterial blood...
When all four heme groups are bound to oxygen, the resulting molecule is called oxyhemoglobin. As a result, arterial blood...
4.5K
Transcytosis of IgG
2.9K
Transcytosis is the process in which molecules are internalized by endocytosis, transported across the cell, and released through exocytosis from the opposite end of the cell. Molecules such as insulin, immunoglobulins, and certain nutrients are transferred through the recycling endosomes by recycling and transcytosis.
IgG molecules from a mother undergo transcytosis starting around 13 weeks of gestation. The amount of IgG transferred and entering the fetal blood circulation increases with...
IgG molecules from a mother undergo transcytosis starting around 13 weeks of gestation. The amount of IgG transferred and entering the fetal blood circulation increases with...
2.9K
Multiple Allele Traits
34.9K
The Concept of Multiple Allelism
34.9K


