PLA2G2E Human

Secreted Phospholipase A2-IIE Human Recombinant
Cat. No.
BT22012
Source
Escherichia Coli.
Synonyms
Group IIE secretory phospholipase A2, EC 3.1.1.4, Phosphatidylcholine 2-acylhydrolase GIIE, GIIE sPLA2, sPLA(2)-IIE, sPLA2-IIE, PLA2G2E.
Appearance
Sterile Filtered lyophilized (freeze-dried) powder.
Purity
Greater than 95% as determined by SDS PAGE.
Usage
Prospec'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.
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Description

Secreted Phospholipase A2-IIE Human Recombinant manufactured with N-terminal His-Tag. PLA2G2E His-Tagged Fusion Protein is 15.8 kDa protein containing 123 amino acid residues of the human secreted phospholipase A2-IIE and 16 additional amino acid residues – His-Tag (underlined).

Product Specs

Introduction
Phospholipase A2 (PLA2) enzymes play a crucial role in hydrolyzing membrane glycerophospholipids. This action releases arachidonic acid (AA), a precursor to eicosanoids like prostaglandins and leukotrienes, which are involved in inflammation and other biological processes. The reaction also produces lysophosholipids, another type of lipid mediator. The secretory PLA2 (sPLA2) family, comprising ten identified isozymes, consists of small, calcium-dependent enzymes secreted outside cells. They are implicated in various biological processes, including eicosanoid production, inflammation, and immune response. Research suggests that this enzyme might contribute to atherosclerosis by hydrolyzing phosphatidylcholine (PC) in lipoproteins within the arterial wall. This process could lead to the buildup of bioactive lipids and modified lipoproteins in atherosclerotic plaques. Studies in mice indicate that sPLA2 expression significantly affects HDL particle size and composition. These studies demonstrate that sPLA2 is necessary for the reduction of plasma HDL cholesterol in response to inflammatory stimuli. Furthermore, introducing bacteria into the bronchi of rats led to surfactant degradation and a change in the ratio of large to small surfactant aggregates, suggesting sPLA2 involvement in these processes.
Description
This product is a recombinant human secreted phospholipase A2-IIE protein with an N-terminal His-Tag. With a molecular weight of 15.8 kDa, this fusion protein consists of 123 amino acid residues from the human secreted phospholipase A2-IIE and an additional 16 amino acid residues for the His-Tag (underlined).
Physical Appearance
This product is provided as a sterile, lyophilized (freeze-dried) powder.
Formulation
This product is sterile filtered and lyophilized from a solution of 0.5 mg/ml in 0.05M Acetate buffer with a pH of 4.
Solubility
To reconstitute the lyophilized protein, add 0.2 ml of 0.1M Acetate buffer (pH 4) to the vial and allow the pellet to dissolve completely. For use at higher pH values, it is recommended to dilute the reconstituted protein with the appropriate buffer to a final concentration of 10 µg/ml. Please note that the solubility of this antigen is limited at higher concentrations.
Stability
Lyophilized protein should be stored at -20°C. After reconstitution, aliquot the protein to prevent repeated freezing and thawing cycles. Reconstituted protein can be stored at 4°C for a short period; no significant changes are observed after two weeks at this temperature.
Purity
The purity of this product is greater than 95% as determined by SDS PAGE.
Synonyms
Group IIE secretory phospholipase A2, EC 3.1.1.4, Phosphatidylcholine 2-acylhydrolase GIIE, GIIE sPLA2, sPLA(2)-IIE, sPLA2-IIE, PLA2G2E.
Source
Escherichia Coli.
Amino Acid Sequence

MRGSHHHHHH GMASHMNLVQ FGVMIEKMTG KSALQYNDYG CYCGIGGSHW PVDQTDWCCH AHDCCYGRLE KLGCEPKLEK YLFSVSERGI FCAGRTTCQR LTCECDKRAA LCFRRNLGTY NRKYAHYPNK LCTGPTPPC

Product Science Overview

Introduction

Secreted Phospholipase A2-IIE (sPLA2-IIE) is a member of the secreted phospholipase A2 (sPLA2) family, which are enzymes involved in the hydrolysis of phospholipids at the sn-2 position, releasing free fatty acids and lysophospholipids. These enzymes play crucial roles in various physiological processes, including inflammation, host defense, and lipid metabolism .

Structure and Function

sPLA2-IIE is characterized by its high content of disulfide bonds, typically 6-8, which contribute to its stability and activity. The enzyme specifically targets the sn-2 acyl bond of phospholipids, leading to the production of arachidonic acid, a precursor for pro-inflammatory eicosanoids . The human recombinant form of sPLA2-IIE is often produced with an N-terminal His-Tag to facilitate purification and study .

Expression and Purification

The recombinant sPLA2-IIE is commonly expressed in the yeast Pichia pastoris, which allows for high-yield production. The protein is purified using a combination of cation exchange and size-exclusion chromatography. This method ensures the production of high-purity sPLA2-IIE, which is essential for functional and structural studies .

Biological Significance

sPLA2-IIE is involved in various biological processes:

  • Inflammation: By releasing arachidonic acid, sPLA2-IIE contributes to the inflammatory response.
  • Host Defense: The enzyme plays a role in the defense against bacterial infections by hydrolyzing bacterial membrane phospholipids.
  • Lipid Metabolism: sPLA2-IIE is involved in the remodeling of cellular membranes and the generation of bioactive lipid mediators .
Research and Applications

The study of recombinant sPLA2-IIE has provided insights into its role in disease mechanisms, particularly in inflammatory and cardiovascular diseases. The high-purity recombinant protein is used in various assays to understand its enzymatic activity, substrate specificity, and potential as a therapeutic target .

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