scholarly journals A double leucine within the GLUT4 glucose transporter COOH-terminal domain functions as an endocytosis signal.

1994 ◽  
Vol 126 (4) ◽  
pp. 979-989 ◽  
Author(s):  
S Corvera ◽  
A Chawla ◽  
R Chakrabarti ◽  
M Joly ◽  
J Buxton ◽  
...  
Keyword(s):  
Cell Surface ◽  
Glucose Transporter ◽  
Surface Display ◽  
Cell Surface Display ◽  
Underlying Mechanism ◽  
Terminal Domain ◽  
Amino Acid Region ◽  
Intracellular Compartments ◽  
A Cell ◽  
Glucose Transporter Glut4

The unique COOH-terminal 30-amino acid region of the adipocyte/skeletal muscle glucose transporter (GLUT4) appears to be a major structural determinant of this protein's perinuclear localization, from where it is redistributed to the cell surface in response to insulin. To test whether an underlying mechanism of this domain's function involves glucose transporter endocytosis rates, transfected cells were generated expressing exofacial hemagglutinin epitope (HA)-tagged erythrocyte/brain glucose transporter (GLUT1) or a chimera containing the COOH-terminal 30 amino acids of GLUT4 substituted onto this GLUT1 construct. Incubation of COS-7 or CHO cells expressing the HA-tagged chimera with anti-HA antibody at 37 degrees resulted in an increased rate of antibody internalization compared to cells expressing similar levels of HA-tagged GLUT1, which displays a cell surface disposition. Colocalization of the internalized anti-HA antibody in vesicular structures with internalized transferrin and with total transporters was established by digital imaging microscopy, suggesting the total cellular pool of transporters are continuously recycling through the coated pit endocytosis pathway. Mutation of the unique double leucines 489 and 490 in the rat GLUT4 COOH-terminal domain to alanines caused the HA-tagged chimera to revert to the slow endocytosis rate and steady-state cell surface display characteristic of GLUT1. These results support the hypothesis that the double leucine motif in the GLUT4 COOH terminus operates as a rapid endocytosis and retention signal in the GLUT4 transporter, causing its localization to intracellular compartments in the absence of insulin.

scholarly journals Insulin Increases Cell Surface GLUT4 Levels by Dose Dependently Discharging GLUT4 into a Cell Surface Recycling Pathway

2004 ◽  
Vol 24 (14) ◽  
pp. 6456-6466 ◽  
Author(s):  
Roland Govers ◽  
Adelle C. F. Coster ◽  
David E. James
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Glucose Transporter ◽  
Insulin Dose ◽  
Cell Types ◽  
Insulin Stimulation ◽  
Intracellular Compartments ◽  
A Cell ◽  
Recycling Pathway ◽  
Glucose Transporter Glut4

ABSTRACT The insulin-responsive glucose transporter GLUT4 plays an essential role in glucose homeostasis. A novel assay was used to study GLUT4 trafficking in 3T3-L1 fibroblasts/preadipocytes and adipocytes. Whereas insulin stimulated GLUT4 translocation to the plasma membrane in both cell types, in nonstimulated fibroblasts GLUT4 readily cycled between endosomes and the plasma membrane, while this was not the case in adipocytes. This efficient retention in basal adipocytes was mediated in part by a C-terminal targeting motif in GLUT4. Insulin caused a sevenfold increase in the amount of GLUT4 molecules present in a trafficking cycle that included the plasma membrane. Strikingly, the magnitude of this increase correlated with the insulin dose, indicating that the insulin-induced appearance of GLUT4 at the plasma membrane cannot be explained solely by a kinetic change in the recycling of a fixed intracellular GLUT4 pool. These data are consistent with a model in which GLUT4 is present in a storage compartment, from where it is released in a graded or quantal manner upon insulin stimulation and in which released GLUT4 continuously cycles between intracellular compartments and the cell surface independently of the nonreleased pool.

scholarly journals Development of a cell surface display system in Chlamydomonas reinhardtii

2022 ◽  
Vol 61 ◽  
pp. 102570
Author(s):  
João Vitor Dutra Molino ◽  
Roberta Carpine ◽  
Karl Gademann ◽  
Stephen Mayfield ◽  
Simon Sieber
Keyword(s):  
Cell Surface ◽  
Chlamydomonas Reinhardtii ◽  
Surface Display ◽  
Cell Surface Display ◽  
Display System ◽  
A Cell ◽  
Cell Surface Display System

scholarly journals Rapidly rendering cells phagocytic through a cell surface display technique and concurrent Rac activation

2014 ◽  
Vol 7 (334) ◽  
pp. rs4-rs4 ◽  
Author(s):  
H. Onuma ◽  
T. Komatsu ◽  
M. Arita ◽  
K. Hanaoka ◽  
T. Ueno ◽  
...  
Keyword(s):  
Cell Surface ◽  
Surface Display ◽  
Cell Surface Display ◽  
A Cell ◽  
Display Technique

scholarly journals A cell surface display fluorescent biosensor for measuring MMP14 activity in real-time

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Alexander Braun ◽  
Matthew J. Farber ◽  
Zachary A. Klase ◽  
Peter B. Berget ◽  
Kenneth A. Myers
Keyword(s):  
Cell Surface ◽  
Real Time ◽  
Surface Display ◽  
Cell Surface Display ◽  
Fluorescent Biosensor ◽  
A Cell

scholarly journals Development of a Cell Surface Display System in a Magnetotactic Bacterium, “Magnetospirillum magneticum” AMB-1

2008 ◽  
Vol 74 (11) ◽  
pp. 3342-3348 ◽  
Author(s):  
Masayoshi Tanaka ◽  
Yuko Nakata ◽  
Tetsushi Mori ◽  
Yoshiko Okamura ◽  
Hitoshi Miyasaka ◽  
...  
Keyword(s):  
Magnetic Nanoparticles ◽  
Cell Surface ◽  
Magnetic Force ◽  
Secondary Structure Prediction ◽  
Surface Display ◽  
Cell Surface Display ◽  
Display System ◽  
A Cell ◽  
Magnetospirillum Magneticum ◽  
Cell Surface Display System

ABSTRACT Bacterial cell surface display is a widely used technology for bioadsorption and for the development of a variety of screening systems. Magnetotactic bacteria are unique species of bacteria due to the presence of magnetic nanoparticles within them. These intracellular, nanosized (50 to 100 nm) magnetic nanoparticles enable the cells to migrate and be manipulated by magnetic force. In this work, using this unique characteristic and based on whole-genomic and comprehensive proteomic analyses of these bacteria, a cell surface display system has been developed by expressing hexahistidine residues within the outer coiled loop of the membrane-specific protein (Msp1) of the “Magnetospirillum magneticum” (proposed name) AMB-1 bacterium. The optimal display site of the hexahistidine residues was successfully identified via secondary structure prediction, immunofluorescence microscopy, and heavy metal binding assay. The established AMB-1 transformant showed high immunofluorescence response, high Cd2+ binding, and high recovery efficiency in comparison to those of the negative control when manipulated by magnetic force.

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