Cell surface glycoproteins of CHO cells. I. Internalization and rapid recycling

Journal Article · Wed Jan 01 04:00:00 EST 1986 · Exp. Cell Res.; (United States)

DOI:https://doi.org/10.1016/0014-4827(86)90534-3· OSTI ID:5783694

Raub, T J; Denny, J B; Roberts, R M

The major cell surface proteins of Chinese hamster ovary (CHO) cells have been investigated after reacting cells at 4/sup 0/C with the membrane-impermeant reagent, trinitrobenzenesulfonate (TNBS). Immunoprecipitation and subsequent two-dimensional, sodiumdodecyl sulfate, polyacrylamide gel electrophoresis (SDS-PAGE) of proteins from derivatized cells that had been labelled previously with (/sup 3/H)D-glucosamine or (/sup 3/H)L-leucine showed that TNBS reacted with most of the high molecular weight (HMW) acidic glycoproteins that became labelled with iodine by the lactoperoxidase technique and that bind the lectin, wheat germ agglutinin (WGA). After warming the cells to allow endocytosis to proceed, molecule haptenized with trinitrophenol (TNP) groups were followed radio-chemically by means of (/sup 125/I)anti-DNP antibodies. Within 15 min at 37/sup 0/C, a steady-state between surface and cytoplasmic label was reached, with about 65% of the hapten located internally. Recycling of internalized TNP groups back to the cell surface also occurred rapidly (t/sub 1/2/ approx. 5 min). Our results are consistent with the view that the majority of plasma membrane glycoproteins are continuously being internalized and recycled at a high rate.

• Research Organization: Univ. of Florida, Gainesville
• OSTI ID: 5783694
• Journal Information: Exp. Cell Res.; (United States), Journal Name: Exp. Cell Res.; (United States) Vol. 165; ISSN ECREA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
550301 -- Cytology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
AMINO ACIDS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
CARBOHYDRATES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CELL MEMBRANES
CELL WALL
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
GLUCOPROTEINS
GLUCOSAMINE
HAMSTERS
HEXOSAMINES
HEXOSES
IMMUNOASSAY
IMMUNOLOGY
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LEUCINE
MAMMALS
MEMBRANES
MONOCLONAL ANTIBODIES
MONOSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNOLOGY
RADIOISOTOPES
RODENTS
SACCHARIDES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES