Recombinant Human NHEJ1 Protein, N-His

Description of Recombinant Human NHEJ1 Protein, N-His

Introduction

The NHEJ1 protein, also known as DNA-PKcs, is a key component of the non-homologous end joining (NHEJ) pathway, which is responsible for repairing DNA double-strand breaks (DSBs). Recombinant human NHEJ1 protein, produced through genetic engineering techniques, has become an essential tool in the study of DNA repair mechanisms and has potential applications in cancer therapy and gene therapy. In this article, we will explore the structure, activity, and applications of recombinant human NHEJ1 protein.

Structure of Recombinant Human NHEJ1 Protein

The NHEJ1 protein is a large polypeptide chain consisting of 4128 amino acids with a molecular weight of approximately 469 kDa. It is composed of multiple domains, including the N-terminal domain, the kinase domain, the helical domain, and the C-terminal domain. The N-terminal domain is responsible for binding to DNA and other proteins, while the kinase domain is essential for the catalytic activity of the protein. The helical domain plays a crucial role in the interaction between NHEJ1 and other proteins involved in the NHEJ pathway, and the C-terminal domain is involved in the regulation of the protein’s activity.

Recombinant human NHEJ1 protein is produced by expressing the NHEJ1 gene in a suitable host cell, such as E. coli or mammalian cells. The protein is then purified using various techniques, such as chromatography and gel filtration, to obtain a highly pure and active form of the protein.

Activity of Recombinant Human NHEJ1 Protein

The main function of NHEJ1 protein is to repair DNA DSBs, which can be caused by various factors, such as radiation, chemicals, and errors during DNA replication. The NHEJ pathway is the primary mechanism for repairing these breaks in mammalian cells, and NHEJ1 is a crucial component of this pathway.

Upon binding to a DSB, NHEJ1 recruits other proteins, including Ku70/80 and DNA ligase IV, to form a complex that repairs the break. The kinase domain of NHEJ1 plays a crucial role in this process by phosphorylating other proteins, which helps in the recruitment and activation of other repair enzymes. Additionally, the helical domain of NHEJ1 is involved in the proper alignment of the broken DNA ends, ensuring accurate repair.

Recombinant human NHEJ1 protein has been extensively studied in vitro, and its activity has been shown to be similar to that of the native protein. It has also been successfully used in reconstitution experiments to study the NHEJ pathway and its components.

Applications of Recombinant Human NHEJ1 Protein

Recombinant human NHEJ1 protein has various applications in both research and medicine. Its role in DNA repair makes it a valuable tool for studying the mechanisms of DNA damage and repair. It has been used in various in vitro and in vivo experiments to understand the role of NHEJ1 in different cellular processes, such as cell cycle regulation and genomic stability.

Moreover, NHEJ1 protein has potential applications in cancer therapy. Cancer cells often have defects in the NHEJ pathway, making them more dependent on other DNA repair mechanisms. Inhibiting NHEJ1 activity in these cells can lead to the accumulation of DNA damage, resulting in cell death. Recombinant NHEJ1 protein can be used to study this potential therapeutic approach and develop targeted cancer treatments.

Another potential application of recombinant NHEJ1 protein is in gene therapy. The NHEJ pathway is essential for the repair of DNA DSBs caused by gene editing techniques such as CRISPR-Cas9. The use of recombinant NHEJ1 protein can enhance the efficiency of gene editing by promoting accurate repair of these breaks