HIV-1 Integrase

HIV-1 Integrase Recombinant
Cat. No.
BT21822
Source
Escherichia Coli.
Synonyms
Appearance
Sterile filtered colorless clear solution.
Purity
Greater than 95.0% as determined by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
Shipped with Ice Packs
In Stock

Description

The E.coli derived 36 kDa recombinant protein is a non-glycosylated polypeptide chain, containing the HIV-1 immunodominant regions from the pol protein (intergrase) and fused with a six histidines tag.

Product Specs

Introduction
Integrase, an enzyme produced by HIV, enables the integration of its genetic material into the host cell's DNA. This enzyme, a crucial part of the pre-integration complex, consists of three domains: an N-terminal HH-CC zinc finger domain (partially responsible for multimerization), a central catalytic domain, and a C-terminal domain. Both the central catalytic and C-terminal domains exhibit binding affinity for viral and cellular DNA. Currently, there are no crystal structures available that depict Integrase bound to its DNA substrates. HIV-1 integrase functions in dimeric or tetrameric forms. Furthermore, several host cell proteins interact with and potentially facilitate the integration process.
Description
This 36 kDa recombinant protein, derived from E. coli, is a non-glycosylated polypeptide chain. It encompasses the immunodominant regions of the HIV-1 pol protein (integrase) and is fused with a six-histidine tag.
Physical Appearance
The product is a sterile, filtered solution that is colorless and clear.
Formulation
The protein is supplied in a solution containing 1.5M urea, 25mM Tris-HCl (pH 8.0), 0.2% Triton-X, and 50% glycerol.
Purity
The purity of the HIV-1 Integrase is greater than 95% as determined by SDS-PAGE analysis.
Stability
While HIV-1 Integrase remains stable for one week at 4°C, it is recommended to store the protein below -18°C to ensure long-term stability. Repeated freezing and thawing should be avoided.
Applications
HIV-1 Integrase antigen is suitable for use in ELISA and Western blot assays. It serves as an excellent antigen for the early detection of HIV seroconversion, exhibiting minimal specificity issues.
Source
Escherichia Coli.
Amino Acid Sequence

MFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVASCDKCQLKGEA
MHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQETAYFILK
LAGRWPVKTIHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMN
KELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTK
ELQKQITKIQNFRVYYRDSRDPLWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKA
KIIRDYGKQMAGDDCVASRQDEDHHHHHH.

Specificity
Immunoreactive with all sera of HIV-1 infected individuals.

Product Science Overview

Introduction

Human Immunodeficiency Virus type 1 (HIV-1) integrase is a crucial enzyme in the life cycle of HIV. It facilitates the integration of viral DNA into the host cell genome, a critical step for viral replication and persistence. Recombinant HIV-1 integrase is a form of this enzyme produced through recombinant DNA technology, allowing for detailed study and drug development.

Structure and Function

HIV-1 integrase is composed of three domains:

  1. N-terminal zinc-binding domain (NTD): Involved in multimerization of the enzyme.
  2. Catalytic core domain (CCD): Contains the active site and is responsible for the catalytic activity of the enzyme.
  3. C-terminal DNA-binding domain (CTD): Binds to viral DNA and is essential for integration.

The enzyme catalyzes two key reactions:

  1. 3’-processing: Cleavage of a dinucleotide from each 3’ end of the viral DNA.
  2. Strand transfer: Insertion of the processed viral DNA into the host genome.
Recombinant Production

Recombinant HIV-1 integrase is produced by cloning the integrase gene into an expression vector, which is then introduced into a host cell (commonly E. coli). The host cells express the integrase protein, which can be purified for research purposes. This recombinant protein is used in various assays to study the enzyme’s activity and to screen for potential inhibitors.

Importance in Drug Development

Integrase inhibitors are a class of antiretroviral drugs that target the integrase enzyme, preventing the integration of viral DNA into the host genome. These inhibitors are crucial in the treatment of HIV/AIDS. The development of recombinant HIV-1 integrase has been instrumental in the discovery and optimization of these drugs. Early integrase inhibitors, such as raltegravir, have shown significant efficacy in reducing viral loads in patients .

Research Applications

Recombinant HIV-1 integrase is used in various research applications, including:

  • Structural studies: Understanding the enzyme’s structure and its interaction with DNA and inhibitors.
  • Mechanistic studies: Investigating the detailed mechanisms of 3’-processing and strand transfer.
  • Drug screening: Identifying and optimizing new integrase inhibitors.
Challenges and Future Directions

Despite the success of integrase inhibitors, resistance mutations can arise, reducing the efficacy of these drugs. Research continues to focus on understanding the mechanisms of resistance and developing next-generation inhibitors that can overcome these challenges .

In conclusion, recombinant HIV-1 integrase is a vital tool in the fight against HIV/AIDS, providing insights into the enzyme’s function and aiding in the development of effective antiretroviral therapies.

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