Membranes of Animal Cells

Warren, Leonard; Glick, Mary Catherine

doi:10.1083/jcb.37.3.729

Cited by 233 publications

(58 citation statements)

References 46 publications

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“…Studies of the metabolism of the surface membrane molecules have shown a continuous cell surface turnover and release of substances into the surrounding medium (Warren and Glick, 1968). Our studies appear to indicate that the local immune reactions increase this cell surface turnover and subsequent release.…”

Section: Discussionsupporting

confidence: 55%

The Presence of Tumour Specific Membrane Antigen in the Serum of Rats with Chemically Induced Sarcomata

Thomson¹,

Steele²,

Alexander³

1973

Br J Cancer

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Summary.-Antibodies to the tumour-specific transplantation type antigen (TSTA) of a transplanted methylcholanthrene-induced sarcoma (MC-1) in syngeneic rats were studied using the techniques of indirect membrane immunofluorescence and mixed haemadsorption with a 5'Cr-labelled indicator cell. After tumour excision, anti-TSTA antibody was readily measurable in both serum and lymph. In contrast, the tumour-bearing animal had no measurable anti-TSTA antibody in the serum but low titres in the lymph. Consequently, we formed the hypothesis that in the presence of a growing tumour the serum contained antigen-antibody complexes with antigen in excess.To test this hypothesis, tumour-bearing serum was examined for the presence of free antigen and antigen-antibody complexes by 2 different methods. In the first method, tumour-bearing serum was cross-linked with glutaraldehyde and was found to absorb specifically the anti-TSTA antibody, indicating free circulating TSTA. Next, antigen-antibody complexes were split with salt or acid and separated into a low molecular weight (or " antigen ") fraction (<100,000) and a high molecular weighit (or "antibody") fraction (>100,000). The low M.W. fraction specifically inhibited the anti-TSTA antibody when tested by either membrane immunofluorescence or mixed haemadsorption, indicating the presence of antigen from antigen-antibody complexes in the tumour-bearing circulation. The possible effect on the host's immune response of circulating free tumour antigen and antigenantibody complexes are discussed.

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Section: Discussionsupporting

confidence: 55%

The Presence of Tumour Specific Membrane Antigen in the Serum of Rats with Chemically Induced Sarcomata

Thomson¹,

Steele²,

Alexander³

1973

Br J Cancer

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“…L-cell membrane turnover was studied using a number of different internal labels, [l~C]leucine, [l'C]glucosamine, and [~4C]glucose (33). The last label is rather nonspecific and no attempt was made to identify the species labeled.…”

Section: Other Studies Of Membrane Protein Turnovermentioning

confidence: 99%

“…Different techniques have been used (3,4,8,17,20,25,33) but most suffer from the necessity of isolating membranes of varying purity and yield as well as the problem of reutilization of label. Use of the enzymatic iodination technique to label only the externally disposed membrane proteins provided a reasonable solution to both problems.…”

mentioning

confidence: 99%

Externally disposed plasma membrane proteins. II. Metabolic fate of iodinated polypeptides of mouse L cells.

Hubbard¹,

Cohn²

1975

The Journal of Cell Biology

137

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The fate of the L-cell plasma membrane proteins labeled by enzymatic iodination was studied. The disappearance of label from growing cells exhibits a biphasic behavior, with 5-20% lost rapidly (t~ ~ 2 h) and 80-90% lost relatively slowly (t~ ~ 25-33 h). The loss is temperature dependent and serum independent, and is accompanied by the appearance of 51% [l~5l]monoiodotyrosine (MIT) in the medium by 47 h. A variable amount (1-14%) of acid-insoluble label can be recovered in the medium over 47 h. Sodium dodecyl sulfate (SDS)-polyacrylamide gel labeling patterns from cells cultured up to 48 h after iodination reveal no change in the relative distribution of radioactivity, indicating similar rates of degradation for most of the labeled membrane proteins.The fate of the labeled membrane proteins was studied at various times after phagocytosis of nondigestible polystyrene particles. Iodinated L cells phagocytose sufficient 1.1 /~m latex beads in 60 min to interiorize 15-30% of the total cell surface area. Electron microscope autoradiography confirmed that labeled membrane is internalized during phagocytosis. The latex-containing phagocytic vacuoles are isolated by flotation in a discontinuous sucrose gradient. 15-30% of the total incorporated label and a comparable percentage of alkaline phosphodiesterase I activity (PDase, a plasma membrane enzyme marker) are recovered in the phagocytic vacuole fraction. Lysosomal enzyme activities are found in the latex vacuole fraction, indicating formation of phagolysosomes. SDS gel analyses reveal that all of the radioactive proteins initially present on the intact cell's surface are interiorized to the same relative extent. Incorporated label and PDase activity disappear much more rapidly from the phagolysosomes than from the whole cell. In the phagolysosomal compartment, >70% of the TCA-precipitable labeled proteins and all of the PDase activity are lost rapidly (tv, = 1-2 h) but ~ 30% of the labeled proteins in this compartment are degraded with a 17-20-h half-life. The slowly degraded label is due to specific long-lived polypeptides, of 85,000 and 8,000--15,000 daltons, which remain in the phagolysosomal membrane up to 40 h after phagocytosis.

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“…Likewise, we do not know whether the other membrane proteins are also turning over, but at a rate too slow to be seen in our experiments. It should be noted that there is extensive turnover of membranes in cultured mammalian cells, even when they are in a stationary, nongrowing state (36).…”

Section: Proteins Synthesized By Intact Reticulocytesmentioning

confidence: 99%

Biosynthesis of Reticulocyte Membrane Proteins by Membrane-Free Polyribosomes

Lodish

1973

Proc. Natl. Acad. Sci. U.S.A.

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Intact rabbit reticulocyte cells synthesize two predominant species of membrane polypeptides; at least 10 reticulocyte membrane polypeptide species are not produced by the cells. Cell-free extracts of reticulocytes, free of any membranes or membrane-bound polyribosomes, synthesize large amounts of these two membrane polypeptides; one of these polypeptides appears to be modified, probably by loss of 20-40 amino acids, after it is incorporated into the membrane. Deprivation of hemin results in inhibition of synthesis by lysates of membrane proteins, globin, and all other cytoplasmic proteins.The protein composition of mammalian erythrocyte membrane is now fairly well established. There exist 7-9 main components (1-8); the major species include two polypeptides of molecular weight greater than 200,000 (4), a protein of molecular weight 100,000, and a sialoylgylcoprotein which contains the MN, A, and B blood-group activities (8). The latter two species are the only membrane proteins found on the cell surface (3, 9-12); they may also penetrate to the interior surface of the membrane (13-15). Little is known, however, about the biosynthesis of erythrocyte membrane proteins: the types of erythropoetic cells that synthesize each protein, the type of polysome (free or membrane-attached) that synthesizes the proteins, and the factors that regulate their synthesis.The reticulocyte is the penultimate cell in the erythropoetic series; it has no nucleus, and makes no RNA or DNA.About 90% of protein produced is a and ,3-globin chains (16,17). The protein composition of reticulocyte membranes is very similar to that of erythrocyte membranes (my unpublished observations). In this paper, I show that reticulocytes synthesize only 2 major and 2-3 minor membrane proteins; most of the membrane proteins are no longer being synthesized in significant amounts at this stage.In most mammalian cells a fraction of the polyribosomes are firmly attached to membranes; these are the site of synthesis of most, if not all, proteins that are eventually transported out of the cell, such as trypsinogen, chymotrypsinogen, albumin, and other serum proteins (18-22). It is not at all clear that this is the sole function of these polysomes.Burka has claimed that rabbit reticulocytes contain membrane-bound polyribosomes (23) that synthesize most of the nonglobin proteins made by the cell (24). By contrast, we showed recently that membrane-free reticulocyte polyribosomes will produce precisely the same globin and nonglobin proteins as does the intact cell (17). In this paper I show, using dodecyl sulfate-gel electrophoresis and peptide map-1526 ping procedures, that a membrane-free lysate makes large amounts of the two membrane polypeptides produced by the intact cell; one of these proteins appears to be modified, probably by removal of 20-40 amino acids when it is incorporated into the membrane. MATERIALS AND METHODSMaterials. Acetylphenylhydrazine was purchased from Sigma, and [35S]methionine (30,000 Ci/mol) from Amersham/Searle. Sources of all o...

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Membranes of Animal Cells

Cited by 233 publications

References 46 publications

The Presence of Tumour Specific Membrane Antigen in the Serum of Rats with Chemically Induced Sarcomata

The Presence of Tumour Specific Membrane Antigen in the Serum of Rats with Chemically Induced Sarcomata

Externally disposed plasma membrane proteins. II. Metabolic fate of iodinated polypeptides of mouse L cells.

Biosynthesis of Reticulocyte Membrane Proteins by Membrane-Free Polyribosomes

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