Spectrin Rearrangement Early in Erythrocyte Ghost Endocytosis

The endocytic vacuoles induced in white ghosts were found to be depleted of spectrin and therefore it was proposed that they arose from spectrin-free areas in the erythrocyte membrane. To follow changes in spectrin distribution during endocytosis, affinity-purified rabbit antispectrin antibodies wer...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of cell biology 1979-09, Vol.82 (3), p.654-663
Main Authors: Hardy, Britta, Bensch, K. G., Schrier, S. L.
Format: Article
Language:English
Subjects:
Adenosine Triphosphate - pharmacology
Alkaline Phosphatase - pharmacology
Antibodies
Binding Sites, Antibody
Cell membranes
Edetic Acid - pharmacology
Endocytosis
Endocytosis - drug effects
Erythrocyte membrane
Erythrocyte Membrane - metabolism
Erythrocytes
Erythrocytes - metabolism
Ferritins
Humans
Membrane Proteins - metabolism
Phosphatases
Spectral methods
Spectrin - immunology
Spectrin - metabolism
Spectroscopic analysis
Trypsin - pharmacology
Vacuoles
Vacuoles - metabolism
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The endocytic vacuoles induced in white ghosts were found to be depleted of spectrin and therefore it was proposed that they arose from spectrin-free areas in the erythrocyte membrane. To follow changes in spectrin distribution during endocytosis, affinity-purified rabbit antispectrin antibodies were produced. Quantitative techniques were developed for the use of a highly specific125I- F( ab′)2antispectrin, and these showed that before the appearance of vacuoles, as assessed by phase microscopy, there was a reproducible decrease in immunoreactive spectrin. To determine whether this spectrin decrease represented a local or diffuse spectrin loss or a spectrin rearrangement, morphologic studies were undertaken using transmission electron microscopy on samples treated with rabbit antispectrin and ferritin-conjugated goat anti-rabbit immunoglobulin. These studies showed that endocytosis was preceded by the creation of extensive spectrin-free areas separated by discrete spectrin-containing zones. Pretreatment of ghosts with alkaline phosphatase blocked all forms of endocytosis and prevented the creation of spectrin-free areas. Therefore, it is proposed that under the impetus of endocytosis inducers, phosphorylated spectrin is redistributed so that spectrin-free zones are created, and that endocytic vacuoles form and fuse in spectrin-free areas.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.82.3.654