sPLA2-IIA Human (E. coli)

Description

Total 140 AA. MW: 15.8 kDa (calculated). N-Terminal His-tag, 16 extra AA (highlighted).

Amino Acid Sequence

Source

E. coli

Purity

>95%

SDS-PAGE gel

12% SDS-PAGE separation of Human sPLA2-IIA 1. M.W. marker – 14, 21, 31, 45, 66, 97 kDa 2. reduced and heated sample, 10μg/lane 3. non-reduced and non-heated sample, 10μg/lane

Formulation

Filtered (0,4 μm) and lyophilized in 0.5 mg/mL in 0.05M Acetate buffer pH4

Reconstitution

Add 0.1M Acetate buffer pH4 to prepare a working stock solution of approximately 0.5 mg/mL and let the lyophilized pellet dissolve completely. For conversion into higher pH value, we recommend intensive dilution by relevant buffer to a concentration of 10μg/mL. In higher concentrations the solubility of this antigen is limited. Product is not sterile! Please filter the product by an appropriate sterile filter before using it in the cell culture.

Storage, Stability/Shelf Life

Store lyophilized protein at –20°C. Lyophilized protein remains stable until the expiry date when stored at –20°C. Aliquot reconstituted protein to avoid repeated freezing/thawing cycles and store at –80°C for long term storage. Reconstituted protein can be stored at 4°C for a limited period of time; it does not show any change after two weeks at 4°C.

Quality Control Test

BCA to determine quantity of the protein.

SDS PAGE to determine purity of the protein.

Applications

Western blotting

Note

This product is intended for research use only.

Introduction to the Molecule

Phospholipase A2 (PLA2) catalyzes the hydrolysis of the sn-2 position of membrane glycerophospho­lipids to liberate arachidonic acid (AA), a precursor of eicosanoids including prostaglandins and leukotrienes. The same reaction also produces lysophosholipids, which represent another class of lipid mediators.

The secretory PLA2 (sPLA2) family, in which 10 isozymes have been identified, consists of low-molecular weight, Ca²⁺-requiring secretory enzymes that have been implicated in a number of biological processes, such as modification of eicosanoid generation, inflammation, and host defense.

This enzyme has been proposed to hydrolyze phosphatidylcholine (PC) in lipoproteins to liberate lyso-PC and free fatty acids in the arterial wall, thereby facilitating the accumulation of bioactive lipids and modified lipoproteins in atherosclerotic fo­ci.

In mice, sPLA2 expression significantly influences HDL particle size and composition and demonstrate that an induction of sPLA2 is required for the decrease in plasma HDL cholesterol in response to inflammatory stimuli. Instillation of bacteria into the bronchi was associated with surfactant degradation and a decrease in large:small ratio of surfactant aggregates in rats.

References to this Product

• Ronkko S, Rekonen P, Kaarniranta K, Puustjarvi T, Terasvirta M, Uusitalo H . Phospholipase A2 in chamber angle of normal eyes and patients with primary open angle glaucoma and exfoliation glaucoma. Mol Vis . Mar 26;13:408-17 (2007)

References

• Koduri RS, Gronroos JO, Laine VJ, Le Calvez C, Lambeau G, Nevalainen TJ, Gelb MH. Bactericidal properties of human and murine groups I, II, V, X, and XII secreted phospholipases A(2). J Biol Chem. 2002 Feb 22;277(8):5849-57.
• Murakami M, Masuda S, Shimbara S, Bezzine S, Lazdunski M, Lambeau G, Gelb MH, Matsukura S, Kokubu F, Adachi M, Kudo I. Cellular arachidonate-releasing function of novel classes of secretory phospholipase A2s (groups III and XII). J Biol Chem. 2003 Mar; 278(12): 10657-67.
• Ishizaki J, Suzuki N, Higashino K, Yokota Y, Ono T, Kawamoto K, Fujii N, Arita H, Hanasaki K. Cloning and characterization of novel mouse and human secretory phospholipase A(2)s. J Biol Chem. 1999 Aug; 274(35):24973-9.
• Ito M, Ishikawa Y, Kiguchi H, Komiyama K, Murakami M, Kudo I, Akasaka Y, Ishii T. Distribution of type V secretory phospholipase A2 expression in human hepatocytes damaged by liver disease. J Gastroenterol Hepatol. 2004 Oct;19(10):1140-9.
• Yagami T, Ueda K, Asakura K, Hayasaki-Kajiwara Y, Nakazato H, Sakaeda T, Hata S, Kuroda T, Takasu N, Hori Y. Group IB secretory phospholipase A2 induces neuronal cell death via apoptosis. J Neurochem. 2002 May; 81(3): 449-61.
• Wooton-Kee CR, Boyanovsky BB, Nasser MS, de Villiers WJ, Webb NR. Group V sPLA2 hydrolysis of low-density lipoprotein results in spontaneous particle aggregation and promotes macrophage foam cell formation. Arterioscler Thromb Vasc Biol. 2004 Apr;24(4):762-7.
• Yagami T, Ueda K, Asakura K, Hata S, Kuroda T, Sakaeda T, Takasu N, Tanaka K, Gemba T, Hori Y. Human group IIA secretory phospholipase A2 induces neuronal cell death via apoptosis. Mol Pharmacol. 2002 Jan; 61(1): 114-26.
• Lindbom J, Ljungman AG, Lindahl M, Tagesson C. Increased gene expression of novel cytosolic and secretory phospholipase A(2) types in human airway epithelial cells induced by tumor necrosis factor-alpha and IFN-gamma. J Interferon Cytokine Res. 2002 Sep; 22(9): 947-55. .
• Attalah HL, Wu Y, Alaoui-El-Azher M, Thouron F, Koumanov K, Wolf C, Brochard L, Harf A, Delclaux C, Touqui L. Induction of type-IIA secretory phospholipase A2 in animal models of acute lung injury. Eur Respir J. 2003 Jun; 21(6): 1040-5.
• Balboa MA, Shirai Y, Gaietta G, Ellisman MH, Balsinde J, Dennis EA. Localization of group V phospholipase A2 in caveolin-enriched granules in activated P388D1 macrophage-like cells. J Biol Chem. 2003 Nov 28;278(48):48059-65.
• Touqui L, Alaoui-El-Azher M. Mammalian secreted phospholipases A2 and their pathophysiological significance in inflammatory diseases. Curr Mol Med. 2001 Dec; 1(6): 739-54. Review.
• Murakami M, Kudo I. New phospholipase A(2) isozymes with a potential role in atherosclerosis. Curr Opin Lipidol. 2003 Oct; 14(5): 431-6.
• Valentin E, Ghomashchi F, Gelb MH, Lazdunski M, Lambeau G. On the diversity of secreted phospholipases A(2). Cloning, tissue distribution, and functional expression of two novel mouse group II enzymes. **Cloning, tissue distribution, and functional expression of two novel mouse group II enzymes.J Biol Chem. 1999 Oct; 274(44): 31195-202. Erratum in: J Biol Chem 2000 Jan 275(3): 2246.
• Hanasaki K, Arita H. Phospholipase A2 receptor: a regulator of biological functions of secretory phospholipase A2. Prostaglandins Other Lipid Mediat. 2002 Aug; 68-69: 71-82. Review.
• Silliman CC, Moore EE, Zallen G, Gonzalez R, Johnson JL, Elzi DJ, Meng X, Hanasaki K, Ishizaki J, Arita H, Ao L, England KM, Banerjee A. Presence of the M-type sPLA(2) receptor on neutrophils and its role in elastase release and adhesion. Am J Physiol Cell Physiol. 2002 Oct; 283(4): C1102-13.
• Sawada H, Murakami M, Enomoto A, Shimbara S, Kudo I. Regulation of type V phospholipase A2 expression and function by proinflammatory stimuli. Eur J Biochem. 1999 Aug;263(3):826-35.
• Satake Y, Diaz BL, Balestrieri B, Lam BK, Kanaoka Y, Grusby MJ, Arm JP. Role of group V phospholipase A2 in zymosan-induced eicosanoid generation and vascular permeability revealed by targeted gene disruption. J Biol Chem. 2004 Apr 16;279(16):16488-94.
• Kuwata H, Sawada H, Murakami M, Kudo I. Role of type IIA secretory phospholipase A2 in arachidonic acid metabolism. Adv Exp Med Biol. 1999; 469: 183-8.
• Beck GCh, Yard BA, Schulte J, Haak M, van Ackern K, van der Woude FJ, Kaszkin M. Secreted phospholipases A2 induce the expression of chemokines in microvascular endothelium. Biochem Biophys Res Commun. 2003 Jan 17; 300(3): 731-7.
• Juhl K, Efanov AM, Olsen HL, Gromada J. Secretory phospholipase A2 is released from pancreatic beta-cells and stimulates insulin secretion via inhibition of ATP-dependent K+ channels. Biochem Biophys Res Commun. 2003 Oct 17; 310(2): 274-9.