The Insulin-like Growth Factor II/Mannose 6-Phosphate Receptor Utilizes the Same Membrane Compartments as GLUT4 for Insulin-dependent Trafficking to and from the Rat Adipocyte Cell Surface

Kandror, Konstantin V.; Pilch, Paul F.

doi:10.1074/jbc.271.36.21703

Cited by 56 publications

(56 citation statements)

References 56 publications

(31 reference statements)

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“…This recycling has been shown to involve, at least in part, clathrin-coated pits [12,13] via a dynamin\amphiphysin-dependent mechanism [14], and is consistent with the observed localization of Glut4 to clathrin lattices in the plasma membrane and the partial overlap of Glut4 with the transferrin receptor (TfR) itinerary [15][16][17]. After insulin withdrawal, Glut4 is rapidly internalized from the plasma membrane and is effectively sequestered within the cell until such time as insulin is re-introduced [18][19][20].…”

Section: Introductionsupporting

confidence: 71%

Trafficking of Glut4–Green Fluorescent Protein chimaeras in 3T3-L1 adipocytes suggests distinct internalization mechanisms regulating cell surface Glut4 levels

Campbell

Tavaré

Leader

et al. 1999

Biochemical Journal

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Insulin stimulates glucose transport in adipose and muscle tissue by stimulating the movement (‘translocation’) of an intracellular pool of glucose transporters (the Glut4 isoform) to the plasma membrane. We have engineered a series of chimaeras between Glut4 and green fluorescent protein (GFP) from Aequoria victoria and expressed these proteins in 3T3-L1 adipocytes by microinjection of plasmid cDNA. In the absence of insulin, GFP-Glut4 is localized intracellularly within a perinuclear compartment and multiple intracellular punctate structures. In response to insulin, chimaeric GFP-Glut4 species exhibit a profound redistribution to the cell surface with kinetics comparable with the endogenous protein. The intracellular localization of GFP-Glut4 overlaps partially with compartments labelled with Texas Red transferrin, but is largely distinct from intracellular structures identified using Lysotracker-Red®. K+-depletion resulted in the accumulation of GFP-Glut4 at the cell surface, but to an lesser extent than that observed in response to insulin. In contrast with native Glut4, removal of the insulin stimulus or treatment of insulin-stimulated cells with phosphatidylinositol 3′-kinase inhibitors did not result in re-internalization of the chimaeric GFP-Glut4 from the plasma membrane, suggesting that the recycling properties of this species differ from the native Glut4 molecule. We suggest that the recycling pathway utilized by GFP-Glut4 in the absence of insulin is distinct from that used to internalize GFP-Glut4 from the plasma membrane after withdrawal of the insulin stimulus, which may reflect distinct pathways for internalization of endogenous Glut4 in the presence or absence of insulin.

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Section: Introductionsupporting

confidence: 71%

Trafficking of Glut4–Green Fluorescent Protein chimaeras in 3T3-L1 adipocytes suggests distinct internalization mechanisms regulating cell surface Glut4 levels

Campbell

Tavaré

Leader

et al. 1999

Biochemical Journal

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“…cytes, the protocol we have previously used with IRAP (gp160) (17) and the IGF-II/Man-6-P receptor (8). The most likely reason for this is that our antisera give a high nonspecific background and a low specific signal when used with crude membrane fractions (data not shown), whereas when we use affinity purified GLUT4 vesicles (Fig.…”

Section: Sortilin Is a Major Component Of Glut4-containing Vesicles 2mentioning

confidence: 84%

“…The first two are the IGF-II/Man-6-phosphate receptor (8) and IRAP, respectively (6,7,17). To identify gp110, we isolated adipocytes from 60 rats, prepared the light microsomal fraction from these cells (see "Materials and Methods") and immunoadsorbed Glut4-containing vesicles.…”

Section: Resultsmentioning

confidence: 99%

“…Recently, we were able to detect an additional protein, gp180, which follows the same trafficking pathway as the former proteins but represents a relatively minor component of Glut4 vesicles. We and others have studied these proteins and have identified gp160 as a novel insulin-regulated aminopeptidase, or IRAP (6, 7), gp230 as the IGF-II/Man-6-P receptor (8), and gp180 as the transferrin receptor. 2 Here, we report the identification of gp110, apparently the last major component protein of Glut4-containing vesicles, as the recently described putative sorting protein/ receptor, sortilin (9).…”

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confidence: 99%

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Sortilin Is a Major Protein Component of Glut4-containing Vesicles

Lin

Pilch

Kandror

1997

Journal of Biological Chemistry

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104

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In fat and skeletal muscle cells, glucose transporter isoform 4 (Glut4) is translocated to the cell surface in response to insulin via a system of specialized recycling vesicles. Besides Glut4, these vesicles include the novel insulin-regulatable aminopeptidase, receptors for insulin-like growth factor-II/Man-6-phosphate and transferrin, and a glycoprotein with the molecular mass of 110 kDa. We report here by the criteria of the partial protein sequencing and subsequent cDNA cloning that glycoprotein 110, the last unidentified major protein component of Glut4-containing vesicles, is sortilin, a novel type I receptor-like protein recently cloned from human brain (Petersen, C. M., Nielsen, M. S., Nykjar, A., Jacobsen, L., Tommerup, N., Rasmussen, H. H., Roigaard, H., Gliemann, J., Madsen, P., and Moestrup, S. K. (1997) J. Biol. Chem. 272, 3599 -3605). This protein is highly expressed in fat, brain, and lung and is dramatically up-regulated during differentiation of adipocytes in vitro.The regulation of blood glucose levels by insulin in mammals is achieved by the hormone-dependent movement of the fat and muscle-specific glucose transporter, Glut4, from an intracellular storage vesicle to the cell surface (1-4). As an approach to understand the mechanisms underlying this process, we and others have isolated these vesicles using anti-Glut4 antibodies and have analyzed their protein content by several techniques. Thus, immunoisolation of Glut4-containing vesicles following cell surface biotinylation in the presence of insulin revealed three major component proteins in these vesicles (gp230, gp160, and gp110) 1 that corresponded to major silver staining vesicular proteins present in the basal state (no insulin) (5). These proteins bind to wheat germ agglutinin-agarose and can be detected in an overlay assay with labeled wheat germ lectin, and therefore they are glycoproteins (5). Recently, we were able to detect an additional protein, gp180, which follows the same trafficking pathway as the former proteins but represents a relatively minor component of Glut4 vesicles. We and others have studied these proteins and have identified gp160 as a novel insulin-regulated aminopeptidase, or IRAP (6, 7), gp230 as the IGF-II/Man-6-P receptor (8), and gp180 as the transferrin receptor. 2 Here, we report the identification of gp110, apparently the last major component protein of Glut4-containing vesicles, as the recently described putative sorting protein/ receptor, sortilin (9). MATERIALS AND METHODS Adipocyte Fractionation and Isolation of Glut4-containing Vesicles-Adipocytes were isolated from the epididymal fat pads of male SpragueDawley rats (150 -175 g) by collagenase digestion and fractionated into subcellular fractions by differential centrifugation according to Simpson et al. (10). Light microsomes were immunoadsorbed on monoclonal anti-Glut4 antibody, 1F8 (11), and covalently immobilized on Reacti Gel GF 2000 (Pierce), and the bound material was eluted with 1% Triton X-100 in phosphate-buffered saline. Nonspecific adsorp...

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“…These protocols have resulted in the identification of numerous proteins such as VAMP2 (Cain et al, 1992) and sorting receptors such as M6PR (Kandror and Pilch, 1996) and sortillin (Lin et al, 1997;Morris et al, 1998) that one might expect to be involved in vesicular traffic. Importantly and as cited in the introduction, IRAP was also identified in this way as marker for GSV trafficking that is apparently more abundant than GLUT4 itself.…”

Section: Discussionmentioning

confidence: 99%

p115 Interacts with the GLUT4 Vesicle Protein, IRAP, and Plays a Critical Role in Insulin-stimulated GLUT4 Translocation

Hosaka

Brooks

Presman

et al. 2005

MBoC

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Insulin-regulated aminopeptidase (IRAP) is an abundant cargo protein of Glut4 storage vesicles (GSVs) that traffics to and from the plasma membrane in response to insulin. We used the amino terminus cytoplasmic domain of IRAP, residues 1-109, as an affinity reagent to identify cytosolic proteins that might be involved in GSV trafficking. In this way, we identified p115, a peripheral membrane protein known to be involved in membrane trafficking. In murine adipocytes, we determined that p115 was localized to the perinuclear region by immunofluorescence and throughout the cell by fractionation. By immunofluorescence, p115 partially colocalizes with GLUT4 and IRAP in the perinuclear region of cultured fat cells. The amino terminus of p115 binds to IRAP and overexpression of a N-terminal construct results in its colocalization with GLUT4 throughout the cell. Insulin-stimulated GLUT4 translocation is completely inhibited under these conditions. Overexpression of p115 C-terminus has no significant effect on GLUT4 distribution and translocation. Finally, expression of the p115 N-terminus construct has no effect on the distribution and trafficking of GLUT1. These data suggest that p115 has an important and specific role in insulin-stimulated Glut4 translocation, probably by way of tethering insulin-sensitive Glut4 vesicles at an as yet unknown intracellular site. INTRODUCTIONInsulin normalizes blood glucose levels by mobilizing the muscle and adipocyte glucose transporter isoform, GLUT4, from intracellular storage vesicles and moving it to the plasma membrane (Simpson et al., 2001;Bryant et al., 2002). Various models of GLUT4 trafficking suggest that GLUT4 must exist in more than one intracellular compartment and the major insulin sensitive pool is localized to a compartment that is distinct from endosomal markers and is commonly referred to as glucose transporter storage vesicles (GSVs), or insulin responsive vesicle (IRVs). Despite the critical function of glucose transport in glucose homeostasis, many of the details by which adipocytes and muscle form a pathway of insulin-sensitive GLUT4 trafficking remain unknown. However, it is virtually certain that the major cargo proteins of GSVs must interact with a number of cytosolic and membrane proteins, such as adaptors and tethers, in order to be properly sorted and regulated by insulin.Insulin-responsive aminopeptidase (IRAP) was identified as an abundant cargo protein associated with GLUT4 vesicles that translocates in response to insulin in a manner seemingly identical to GLUT4 Mastick et al., 1994;Keller et al., 1995;Malide et al., 1997;Martin et al., 1997;Ross et al., 1997). In fact, it is more abundantly expressed in vesicles than the transporter (Kupriyanova et al., 2002). When the cytoplasmic N-terminus of IRAP was microinjected into 3T3-L1 adipocytes, GLUT4 was localized on the plasma membrane even in the basal state (Waters et al., 1997), suggesting IRAP can play a role in GSV movement/targeting. A chimeric protein containing the intracellular domain of IRAP and ...

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The Insulin-like Growth Factor II/Mannose 6-Phosphate Receptor Utilizes the Same Membrane Compartments as GLUT4 for Insulin-dependent Trafficking to and from the Rat Adipocyte Cell Surface

Cited by 56 publications

References 56 publications

Trafficking of Glut4–Green Fluorescent Protein chimaeras in 3T3-L1 adipocytes suggests distinct internalization mechanisms regulating cell surface Glut4 levels

Trafficking of Glut4–Green Fluorescent Protein chimaeras in 3T3-L1 adipocytes suggests distinct internalization mechanisms regulating cell surface Glut4 levels

Sortilin Is a Major Protein Component of Glut4-containing Vesicles

p115 Interacts with the GLUT4 Vesicle Protein, IRAP, and Plays a Critical Role in Insulin-stimulated GLUT4 Translocation

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