Abstract
A lectin with strong cytotoxic effect on human colon cancer HT29 and monkey kidney VERO cells was recently identified from the Australian indigenous mushroom Psathyrella asperospora and named PAL. We herein present its biochemical and structural analysis using a multidisciplinary approach. Glycan arrays revealed binding preference towards N-acetylglucosamine (GlcNAc) and, to a lesser extent, towards sialic acid (Neu5Ac). Submicromolar and millimolar affinity was measured by surface plasmon resonance for GlcNAc and NeuAc, respectively. The structure of PAL was resolved by X-ray crystallography, elucidating both the protein's amino acid sequence as well as the molecular basis rationalizing its binding specificity. Proteins 2017; 85:969-975. © 2016 Wiley Periodicals, Inc.
Keywords:
GlcNAc recognition; beta-propeller; fungal lectins.
© 2017 Wiley Periodicals, Inc.
MeSH terms
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Acetylglucosamine / chemistry*
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Acetylglucosamine / metabolism
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Agaricales / chemistry*
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Animals
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Antineoplastic Agents / chemistry*
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Antineoplastic Agents / isolation & purification
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Antineoplastic Agents / metabolism
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Binding Sites
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Carbocyanines / chemistry
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Chlorocebus aethiops
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Crystallography, X-Ray
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Fluorescent Dyes / chemistry
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Fruiting Bodies, Fungal / chemistry
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Fungal Proteins / chemistry*
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Fungal Proteins / isolation & purification
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Fungal Proteins / metabolism
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HT29 Cells
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Humans
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Lectins / chemistry*
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Lectins / isolation & purification
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Lectins / metabolism
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Microarray Analysis
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N-Acetylneuraminic Acid / chemistry*
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N-Acetylneuraminic Acid / metabolism
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Protein Binding
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Protein Conformation, alpha-Helical
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Protein Conformation, beta-Strand
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Protein Interaction Domains and Motifs
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Staining and Labeling
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Surface Plasmon Resonance
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Vero Cells
Substances
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Alexa Fluor 647
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Antineoplastic Agents
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Carbocyanines
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Fluorescent Dyes
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Fungal Proteins
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Lectins
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N-Acetylneuraminic Acid
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Acetylglucosamine