Cryo-EM structure of human glucose transporter GLUT4

Yuan, Yunbin; Kong, Fang; Xu, Hanwen; Zhu, Angqi; Yan, Nieng; Yan, Chuangye

doi:10.1038/s41467-022-30235-5

Cited by 34 publications

(24 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…GLUT1 activity is saturated at 4 mM, which seems well placed for helping to maintain blood glucose levels around 5 mM 61 . The KM for GLUT4 is around 8.2 mM, which is consistent from the data reported on X. oocytes 62 and more recent counterflow transport in proteoliposomes 32 , but has a low turnover of around 1 s-1 . It is interesting to mention that human GLUT4 has the lowest turnover for glucose despite having a higher KM than GLUT1 or GLUT3, which translates with the lowest specific activity (kcat/KM) for glucose (Fig.…”

Section: Assessing Brain Fraction 7 Lipids For Other Glut Transporterssupporting

confidence: 90%

“…Other GLUT isoforms can be recombinantly expressed in Xenopus oocytes [25][26][27] , which has proven a useful host for studying GLUT kinetics and to assess mutations. Crystal and cryo EM structures of GLUT1 [28][29][30] , GLUT3 29,31 , GLUT4 32 , GLUT5 33 and the plasmodium falciparum hexose transporter homologue (PfHT1) 34 have provided a molecular framework for sugar recognition and transport, which is consistent with the biochemical analysis pre-dating structural information 5 . Interestingly, D-glucose is coordinated almost entirely by residues from the C-terminal domain 31 .…”

Section: Introductionsupporting

confidence: 59%

See 1 more Smart Citation

Establishing mammalian GLUT kinetics and lipid preferences in a reconstituted-liposome system

Saudea

Qureshi

McComas

et al. 2022

Preprint

View full text Add to dashboard Cite

Glucose transporters (GLUTs) are essential for organism-wide glucose homeostasis in mammals. Due to their critical role in cellular growth and metabolism, GLUT dysfunction is associated with numerous diseases, such as diabetes and cancer. Despite structural advances, transport assays using purified GLUT have proven to be difficult to implement, hampering deeper mechanistic studies. Indeed, our understanding of the role of GLUT transporters to whole-body glucose homeostasis is largely derived from the kinetics of 2-deoxy-glucose and not D-glucose, as this sugar is rapidly phosphorylated in the cell. Here, we have optimized a transport assay in liposomes for the fructose-specific isoform GLUT5, and demonstrate how inaccuracies can be generated when assessing the impact of mutations and inhibitors with suboptimal activity. By combining lipidomic analysis with native MS and thermal-shift assays, we replicate the GLUT5 transport activities seen in crude lipids with a small number of synthetic lipids. Subsequently, we show how GLUT5 is only active under a specific range of membrane fluidity, and that human GLUT1-4 prefers a similar lipid composition. Although GLUT3 is designated as the high-affinity glucose transporter, in vitro D-glucose kinetics demonstrates that GLUT1 and GLUT3 actually have a similar KM, but GLUT3 has a higher turnover. Interestingly, GLUT4 has a high KM for D-glucose and yet a very slow turnover, which might be to ensure D-glucose uptake is regulated by insulin-dependent trafficking. Overall, our study provides a much-needed transport assay for analyzing in vitro GLUT activities, and implies that high-levels of free fatty acids in membranes, as found in those suffering from metabolic disorders, are a likely contributor to the impairment of glucose transport.

show abstract

Section: Assessing Brain Fraction 7 Lipids For Other Glut Transporterssupporting

confidence: 90%

Section: Introductionsupporting

confidence: 59%

Establishing mammalian GLUT kinetics and lipid preferences in a reconstituted-liposome system

Saudea

Qureshi

McComas

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Inspection of the STP10 structure reveals a unique helical kink in TM10, which is not present in XylE (Figure 4B). An additional structure for inward-open GLUT4 35 was further determined after our initial model training was finalized. Our inward-open GLIUT4 model has a RMSD of 2.3 Å and 1.8 Å before and after MD relaxation, respectively.…”

Section: Generating Sugar Porter Structures For Each Major Conformationmentioning

confidence: 99%

Reconstructing the transport cycle in the sugar porter superfamily using coevolution-powered machine learning

Mitrovic

McComas

Alleva

et al. 2022

Preprint

View full text Add to dashboard Cite

Sugar porters represent the largest group of secondary-active transporters. Some members, such as the glucose (GLUT) transporters, are well-known for their role in maintaining blood glucose homeostasis in mammals, with their expression upregulated in many types of cancers. Because only a few sugar porter structures have been determined, mechanistic models have been constructed by piecing together structural states of distantly-related proteins. Current GLUT transport models are predominantly descriptive, and oversimplified. Here, we have combined coevolution analysis and comparative modeling, to predict structures of the entire sugar porter superfamily in each state of the transport cycle. We have analysed the state-specific contacts inferred from coevolving residue pairs, and shown how this information can be used to rapidly generate free-energy landscapes consistent with experimental estimates, as illustrated here for the mammalian fructose transporter GLUT5. By comparing many different sugar porter models and scrutinizing their sequence, we have been able to define the molecular determinants of the transport cycle, which are conserved throughout the sugar porter superfamily. We have also been able to highlight differences leading to the emergence of proton-coupling, validating and extending the previously proposed latch mechanism. Our computational approach is transferable to any transporter, and to other protein families in general.

show abstract

“…Thus, it is possible that they are more relevant to sensing the low concentrations of monosaccharides generated from starch in the mouth during mastication [ 72 ]. The STR is subject to faster adaptation by endocytosis, as discussed below, while the membrane localization of SGLT and GLUT family members is increased in the plasma membrane (PM) upon glucose stimulation [ 77 , 78 ]. Interestingly, in the enterocytes, mRNA and protein levels and PM localization of SGLT1 are augmented by STR signaling in the neighboring enteroendocrine cells [ 79 ].…”

Section: Pathways Mediating Sweet Taste Signalingmentioning

confidence: 99%

Sweet Taste Signaling: The Core Pathways and Regulatory Mechanisms

Sukumaran

Palayyan

2022

IJMS

View full text Add to dashboard Cite

Sweet taste, a proxy for sugar-derived calories, is an important driver of food intake, and animals have evolved robust molecular and cellular machinery for sweet taste signaling. The overconsumption of sugar-derived calories is a major driver of obesity and other metabolic diseases. A fine-grained appreciation of the dynamic regulation of sweet taste signaling mechanisms will be required for designing novel noncaloric sweeteners with better hedonic and metabolic profiles and improved consumer acceptance. Sweet taste receptor cells express at least two signaling pathways, one mediated by a heterodimeric G-protein coupled receptor encoded by taste 1 receptor members 2 and 3 (TAS1R2 + TAS1R3) genes and another by glucose transporters and the ATP-gated potassium (KATP) channel. Despite these important discoveries, we do not fully understand the mechanisms regulating sweet taste signaling. We will introduce the core components of the above sweet taste signaling pathways and the rationale for having multiple pathways for detecting sweet tastants. We will then highlight the roles of key regulators of the sweet taste signaling pathways, including downstream signal transduction pathway components expressed in sweet taste receptor cells and hormones and other signaling molecules such as leptin and endocannabinoids.

show abstract

Cryo-EM structure of human glucose transporter GLUT4

Cited by 34 publications

References 52 publications

Establishing mammalian GLUT kinetics and lipid preferences in a reconstituted-liposome system

Establishing mammalian GLUT kinetics and lipid preferences in a reconstituted-liposome system

Reconstructing the transport cycle in the sugar porter superfamily using coevolution-powered machine learning

Sweet Taste Signaling: The Core Pathways and Regulatory Mechanisms

Contact Info

Product

Resources

About