Biophysical characterization and structural determination of the potent cytotoxic Psathyrella asperospora lectin.
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.
CERMAV, UPR5301, CNRS and Université Grenoble Alpes, Grenoble, 38041, France.|DCM, UMR5250, CNRS and Université Grenoble Alpes, Grenoble, 38041, France.
João P
JP
Ribeiro
Institute for Glycomics, Griffith University, Gold Coast Campus, Queensland 4222, Australia.
Mohamed
M
Ali Abol Hassan
Institute for Glycomics, Griffith University, Gold Coast Campus, Queensland 4222, Australia.
Razina
R
Rouf
School of Pharmacy and Griffith Health Institute, Griffith University, Gold Coast Campus, Queensland 4222, Australia.
Evelin
E
Tiralongo
Royal Botanic Gardens Victoria, Melbourne, Victoria, 3004, Australia.
Tom W
TW
May
Institute for Glycomics, Griffith University, Gold Coast Campus, Queensland 4222, Australia.
Christopher J
CJ
Day
CERMAV, UPR5301, CNRS and Université Grenoble Alpes, Grenoble, 38041, France.
Anne
A
Imberty
Institute for Glycomics, Griffith University, Gold Coast Campus, Queensland 4222, Australia.
Joe
J
Tiralongo
CERMAV, UPR5301, CNRS and Université Grenoble Alpes, Grenoble, 38041, France.
Annabelle
A
Varrot
D000117:Acetylglucosamine / Q000737:chemistry* / Q000378:metabolism; D000363:Agaricales / Q000737:chemistry*; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D001665:Binding Sites; D002232:Carbocyanines / Q000737:chemistry; D002522:Chlorocebus aethiops; D018360:Crystallography, X-Ray; D005456:Fluorescent Dyes / Q000737:chemistry; D048690:Fruiting Bodies, Fungal / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D037102:Lectins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D046228:Microarray Analysis; D019158:N-Acetylneuraminic Acid / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D000072757:Protein Conformation, beta-Strand; D054730:Protein Interaction Domains and Motifs; D013194:Staining and Labeling; D020349:Surface Plasmon Resonance; D014709:Vero Cells
C569686:Alexa Fluor 647; D000970:Antineoplastic Agents; D002232:Carbocyanines; D005456:Fluorescent Dyes; D005656:Fungal Proteins; D037102:Lectins; D019158:N-Acetylneuraminic Acid; D000117:Acetylglucosamine
GlcNAc recognition; beta-propeller; fungal lectins