A Lectin Purified from Blood Red Bracket Mushroom, Pycnoporus sanguineus (Agaricomycetidae), Mycelium Displayed Affinity Toward Bovine Transferrin.
Fungal lectins constitute excellent ligands for development of affinity adsorbents useful in affinity chromatography. In this work, a lectin was purified from Pycnoporus sanguineus (PSL) mycelium using 3 procedures: by affinity chromatography, using magnetic galactosyl-nanoparticles or galactose coupled to Sepharose, and by ionic exchange chromatography (IEC). The highest lectin yield was achieved by IEC (55%); SDS-PAGE of PSL showed 2 bands with molecular mass of 68.7 and 55.2 kDa and IEC displayed 2 bands at pi 5.5 and 5.2. The lectin agglutinates rat erythrocytes, exhibiting broad specificity toward several monosaccharides, including galactose. The agglutination was also inhibited by the glycoproteins fetal calf fetuin, bovine lactoferrin, bovine transferrin, and horseradish peroxidase. The lectin was then used to synthesize an affinity adsorbent (PSL-Sepharose) and the interaction with glycoproteins was evaluated by analyzing their chromatographic behaviors. The strongest interaction with the PSL-derivative was observed with transferrin, although lower interactions were also displayed toward fetuin and lactoferrin. These results indicate that the purified PSL constitutes an interesting ligand for the design of affinity adsorbents to be used (i.e., in glycoprotein purification).
Catedra de Microbiologia, Departamento de Biociencias, Facultad de Quimica, Universidad de la Republica, Montevideo, Uruguay.
Silvana
S
Albores
Instituto de Nanociencia de Aragon, Universidad de Zaragoza, Zaragoza, Espana.
Maria
M
Moros
Catedra de Microbiologia, Departamento de Biociencias, Facultad de Quimica, Universidad de la Republica, Montevideo, Uruguay.
Maria Pia
MP
Cerdeiras
Instituto de Ciencia de Materiales de Aragon, CSIC, Zaragoza, Espana.
Jesus Martinez
JM
de la Fuente
Instituto de Nanociencia de Aragon, Universidad de Zaragoza, Zaragoza, Espana.
Valeria
V
Grazu
Catedra de Bioquimica, Departamento de Biociencias, Facultad de Quimica, Universidad de la Republica, Montevideo, Uruguay.
Laura Franco
LF
Fraguas
D000371:Agglutination / Q000187:drug effects; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004912:Erythrocytes / Q000187:drug effects; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism; D037102:Lectins / Q000302:isolation & purification* / Q000378:metabolism; D008024:Ligands; D008970:Molecular Weight; D025282:Mycelium / Q000737:chemistry; D055494:Pycnoporus / Q000737:chemistry*; D051381:Rats; D014168:Transferrin / Q000378:metabolism*
D005656:Fungal Proteins; D006023:Glycoproteins; D037102:Lectins; D008024:Ligands; D014168:Transferrin; D005690:Galactose