Identification of Lectins from Metastatic Cancer Cells through Magnetic Glyconanoparticles.
Cancer cells can have characteristic carbohydrate binding properties. Previously, it was shown that a highly metastatic melanoma cell line B16F10 bound to galacto-side-functionalized nanoparticles much stronger than the corresponding less metastatic B16F1 cells. To better understand the carbohydrate binding properties of cancer cells, herein, we report the isolation and characterization of endogenous galactose binding proteins from B16F10 cells using magnetic glyconanoparticles. The galactose-coated magnetic glyconanoparticles could bind with lectins present in the cells and be isolated through magnet-mediated separation. Through Western blot and mass spectrometry, the arginine/serine rich splicing factor Sfrs1 was identified as a galactose-selective endogenous lectin, overexpressed in B16F10 cells, compared with B16F1 cells. In addition, galactin-3 was found in higher amounts in B16F10 cells. Finally, the glyconanoparticles exhibited a superior efficiency in lectin isolation, from both protein mixtures and live cells, than the corresponding more traditional microparticles functionalized with carbohydrates. Thus, the magnetic glyconanoparticles present a useful tool for discovery of endogenous lectins, as well as binding partners of lectins, without prior knowledge of protein identities.
Department of Chemistry, Chemistry Building, Room 426, 578 S. Shaw Lane, Michigan State University, East Lansing, MI 48824 (USA).
Herbert W
HW
Kavunja
Department of Biochemistry and Molecular Biology, Biochemistry Building, Room 402, 603 Wilson Road, Michigan State University, East Lansing, MI 48824 (USA).
Patricia G
PG
Voss
Department of Biochemistry and Molecular Biology, Biochemistry Building, Room 402, 603 Wilson Road, Michigan State University, East Lansing, MI 48824 (USA).
John L
JL
Wang
Department of Chemistry, Chemistry Building, Room 426, 578 S. Shaw Lane, Michigan State University, East Lansing, MI 48824 (USA).
Xuefei
X
Huang
cancer; carbohydrates; lectin; magnetic nanoparticles