PA-IL (LecA)

Ca2+-dependent lectin from Pseudomonas aeruginosa recognizing α- and β-galactosylated glycans

Prices

Product number Product Form Package size In stock Lot number Price
GL-006 PA-IL lectin lyophilized 1 mg 4 12159 160 EUR
5 mg 3 12159 720 EUR
5x 1 mg 1 12159 720 EUR
10 mg 1 12159 1360 EUR
bulk orders
Discounts may be applied for bulk orders. Biotinylated or fluorescently labeled (DyLight) variants can be provided upon request. Contact us at contact@4glyco.cz for prices and availability of those products.

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This product is for R&D use only. Not for human or animal use.

Basic information:

Name: PA-IL (LecA, PA-I)
Organism: Pseudomonas aeruginosa
Expression host: Escherichia coli
Tags: no
Molar mass (monomer): 12762.2 Da
Extinction coefficient: 27960 M-1 cm-1
Oligomeric state: tetramer
PDB code: 1OKO (with Gal)

Protein sequence:
AWKGEVLANNEAGQVTSIIYNPGDVITIVAAGWASYGPTQKWGPQGDREHPDQGLICHDAFCGALVMKIGNSGTIPVNTGLFRWVAPNNVQGAITLIYNDVPGTYGNNSGSFSVNIGKDQS

3D models of two protein structures, one colored in blue and the other in green, featuring complex folds and highlighted active sites with small molecule bindings.

Carbohydrate specificity:

PA-IL (LecA, formerly PA-I) lectin is specific towards carbohydrates containing terminal α-galactosides (bound via α1-3, α1-4, and α1-6 linkages), including αGal1–4Gal motif, typical for Gb3/Pk and P1 antigens, and αGal1-3Gal motif, present in human blood group B antigen [1-2]. Despite certain information in the literature, our data demonstrates that PA-IL binds β-galactosides in a solution.

Ion dependency: Ca2+-dependent
Glycan array data: CFG web pages

Stability:

Stable in a variety of buffers with pH 4.5 - 9.5. Since the presence of calcium ions in the binding site is required for lectin activity, adding calcium ions (0.1-0.5 mM CaCl2) to the working buffer is recommended. Avoid EDTA and other chelating agents as they lead to removing Ca2+ ions from the binding site and losing lectin activity. After reconstitution in neutral pH buffers, the protein should be stable in the fridge for weeks. Adding sodium azide (0.02%) is recommended to avoid microbial growth.

Tm = 72 °C (DSC, 20 mM Tris, 150 mM NaCl, 0.1 mM CaCl2, pH 7.5)

Applications and biological effects:

PA-IL can be used in various applications, including lectin blotting, fluorescence microscopy, flow cytometry, and lectin histochemistry. It is also used in commercial lectin microarrays to study glycosylations of proteins and cells.

PA-IL lectin inhibits ciliary beat in human airways and has cytotoxic effects on respiratory epithelial cells [3, 4]. Furthermore, PA-IL binds efficiently to Gb3-expressing cell lines [1], induces membrane invaginations, and selectively binds cardiac non-myocytes, but not atrial or ventricular cardiomyocytes [5]. The PA-IL-CAR chimeric molecule has recently demonstrated target-specific cytotoxicity against carcinoma cells overexpressing Gb3 (e.g., Burkit´s lymphoma-derived cell lines) [6].

References:

  1. Blanchard et al, J Mol Biol, 2008, doi: 10.1016/j.jmb.2008.08.028
  2. Bojar et al, ACS Chem Biol, 2022, doi: 10.1021/acschembio.1c00689
  3. Bajolat-Laudinat et al, Infect Immun, 1994, doi: 10.1128/iai.62.10.4481-4487.1994
  4. Mewe et al, J Laryngol Otol, 2005, doi: 10.1258/0022215054516313
  5. Darkow et al, Front Physiol, 2020, doi: 10.3389/fphys.2020.00457
  6. Meléndez et al, Cell Mol Life Sci, 2022, doi: 10.1007/s00018-022-04524-7

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