Microfilament association of ASGP-2, the concanavalin A-binding glycoprotein of the cell-surface sialomucin complex of 13762 rat mammary ascites tumor cells

Journal Article · Sat Oct 01 00:00:00 EDT 1988 · Experimental Cell Research; (United States)

DOI:https://doi.org/10.1016/0014-4827(88)90392-8· OSTI ID:5267361

Vanderpuye, L A; Carraway, C A.C.; Carraway, K L ^[1]

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1. Univ. of Miami School of Medicine, FL (USA)

Microfilament-associated proteins and membrane-microfilament interactions are being investigated in microvilli isolated from 13762 rat mammary ascites tumor cells. Phalloidin shift analyses on velocity sedimentation gradients of Triton X-100 extracts of ({sup 3}H)-glucosamine-labeled microvilli identified a 120-kDa cell-surface glycoprotein associated with the microvillar microfilament core. The identification was verified by concanavalin A (Con A) blots of one- and two-dimensional (2D) electrophoresis gels of sedimented microfilament cores. By 2D-electrophoresis and lectin analyses the 120-kDa protein appeared to be a fraction of ASGP-2, the major Con A-binding glycoprotein of the sialomucin complex of the 13762 cells. This identity was confirmed by immunoblot analyses using immunoblot-purified anti-ASGP-2 from anti-membrane serum prepared against microvillar membranes. Proteolysis of the microvilli with subtilisin or trypsin resulted in an increase in the amount of ASGP-2 associated with the microfilament cores. Proteolysis of isolated microvillar membranes, which contain actin but not microfilaments, also increased the association of ASGP-2 with a Triton-insoluble, actin-containing membrane fraction. Since the Triton-insoluble membrane residue is enriched in actin-containing transmembrane complex, which contains a different glycoprotein, the authors suggest that the ASGP-2 is binding indirectly via this complex to the microfilament core in the intact microvilli.

• OSTI ID: 5267361
• Journal Information: Experimental Cell Research; (United States), Vol. 178:2; ISSN 0014-4827
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ASCITES TUMOR CELLS
MORPHOLOGY
GLYCOPROTEINS
TWO-DIMENSIONAL ELECTROPHORESIS
ACTIN
HYDROLYSIS
MUCOPROTEINS
SEDIMENTATION
SIALIC ACID
TRYPSIN
ANIMAL CELLS
CARBOHYDRATES
CHEMICAL REACTIONS
DECOMPOSITION
ELECTROPHORESIS
ENZYMES
HYDROLASES
LYSIS
MONOSACCHARIDES
ORGANIC COMPOUNDS
PEPTIDE HYDROLASES
POLYSACCHARIDES
PROTEINS
SACCHARIDES
SERINE PROTEINASES
SOLVOLYSIS
TUMOR CELLS
550201* - Biochemistry- Tracer Techniques