Crystal Structure of the VHS and FYVE Tandem Domains of Hrs, a Protein Involved in Membrane Trafficking and Signal Transduction

Mao, Yuxin; Nickitenko, Alexei; Duan, Xiaoqun; Lloyd, Thomas E.; Wu, Mark N.; Bellen, Hugo J.; Quiocho, Florante A.

doi:10.1016/s0092-8674(00)80680-7

Cited by 166 publications

(142 citation statements)

References 36 publications

Supporting

Mentioning

139

Contrasting

Order By: Relevance

“…The monomeric GGAs are a multidomain protein family implicated in protein trafficking between the Golgi and endosomes. Previous structural analysis shows that the small 18-kDa VHS domain of the Hrs protein consists of three HEAT or ARM repeats (Mao et al, 2000), a protein fold that has been recently found also in the structure of the regulatory subunit H of the V-ATPase (Sagermann et al, 2001). On the basis of the sequence similarity found, we conclude that also ␤-adaptins and ␤-COP are HEAT or ARM repeat-containing proteins.…”

supporting

confidence: 66%

Subunit H of the V-ATPase Involved in Endocytosis Shows Homology to β-Adaptins

Geyer

Fackler

Peterlin

2002

MBoC

View full text Add to dashboard Cite

The vacuolar ATPase (V-ATPase) is a multisubunit enzyme that facilitates the acidification of intracellular compartments in eukaryotic cells and plays an important role in receptor-mediated endocytosis, intracellular trafficking processes, and protein degradation. In this study we show that the C-terminal fragment of 350 residues of the regulatory subunit H (V1H) of the V-ATPase shares structural and functional homologies with the ␤-chains of adaptor protein complexes. Moreover, the fragment is similar to a region in the ␤-subunit of COPI coatomer complexes, which suggests the existence of a shared domain in these three different families of proteins. For ␤-adaptins, this fragment binds to cytoplasmic di-leucine-based sorting motifs such as in HIV-1 Nef that mediate endocytic trafficking. Expression of this fragment in cells blocks the internalization of transmembrane proteins, which depend on di-leucine-based motifs, whereas mutation of the consensus sequence GEY only partly diminishes the recognition of the sorting motif. Based on recent structural analysis, our results suggest that the di-leucine-binding domain consists of a HEAT or ARM repeat protein fold. INTRODUCTIONTargeting of transmembrane proteins to different compartments of the endocytic and late secretory pathways depends largely on sorting signals contained within their cytosolic domains (Letourneur and Klausner, 1992;Mellman, 1996;Kirchhausen et al., 1997). These signals are thought to interact with specific recognition molecules, which are components of membrane-bound transport intermediates (Schmid, 1997;Hirst and Robinson, 1998;Bonifacino and Dell'Angelica, 1999;Kirchhausen, 1999;Marsh and McMahon, 1999). Most internalized proteins contain sorting signals such as the tyrosine-based motif, Yxx , where x is any amino acid and is a bulky hydrophobic side chain (Lobel et al., 1989;Collawn et al., 1990;Boll et al., 1996) or the di-leucine-based motif, LL (Letourneur and Klausner, 1992). For the tyrosine-based motif, the interaction is mediated by the medium chains of adaptor protein (AP) complexes (Ohno et al., 1995). Cocrystallization of the signal binding domain of 2 with two different Yxx peptides provided a structural explanation for this binding specificity (Owen and Evans, 1998).The Nef protein of primate lentiviruses is the only nontransmembrane protein known to traffic via a di-leucinebased motif (Kirchhausen, 1999). It is required for the internalization of CD4 from the cell surface to endosomes and lysosomes (Craig et al., 1998;Greenberg et al., 1998). Several proteins have been proposed to direct the sorting of Nef. They include the ␤-chain of AP-1 complexes (Bresnahan et al., 1998;Greenberg et al., 1998), the ␤-subunit of the COP-I coatomer (Piguet et al., 1999;Janvier et al., 2001), and the regulatory subunit H (V1H) of the vacuolar ATPase (VATPase), previously named NBP1 for Nef binding protein 1 (Lu et al., 1998;Mandic et al., 2001). Although the precise sites of these interactions varied, all protein complexes were reported to bind ...

show abstract

supporting

confidence: 66%

Subunit H of the V-ATPase Involved in Endocytosis Shows Homology to β-Adaptins

Geyer

Fackler

Peterlin

2002

MBoC

View full text Add to dashboard Cite

show abstract

“…The Hrs and Vps27 proteins contain FTFTN and FSLLN sequences in place of the FSVTV sequence of EEA1, and these exposed loops occupy similar conformations (24,25). Moreover, mutations of the MIL residues of EEA1, Hrs, and Vps27p disrupt the localization of these proteins to membranes or decrease membrane affinity, revealing a crucial role for this element in membrane association (29,37).…”

Section: Resultsmentioning

confidence: 99%

“…This zinc-stabilized module is found in 29 human proteins and has been engineered into an intracellular probe having nanomolar PtdIns(3)P affinity (21,22). Three-dimensional structures of the FYVE domains of EEA1 (13,23), Hrs (24), and Vps27 (25) proteins have been solved. Despite the structural similarity, different models of their membrane orientations have been inferred, emphasizing the need for experimentally derived estimates of membrane insertion.…”

mentioning

confidence: 99%

Multivalent Mechanism of Membrane Insertion by the FYVE Domain

Kutateladze

Capelluto

Ferguson

et al. 2004

Journal of Biological Chemistry

109

View full text Add to dashboard Cite

Targeting of a wide variety of proteins to membranes involves specific recognition of phospholipid head groups and insertion into lipid bilayers. For example, proteins that contain FYVE domains are recruited to endosomes through interaction with phosphatidylinositol 3-phosphate (PtdIns(3)P). However, the structural mechanism of membrane docking and insertion by this domain remains unclear. Here, the depth and angle of micelle insertion and the lipid binding properties of the FYVE domain of early endosome antigen 1 are estimated by NMR spectroscopy. Spin label probes incorporated into micelles identify a hydrophobic protuberance that inserts into the micelle core and is surrounded by interfacially active polar residues. A novel proxyl PtdIns(3)P derivative is developed to map the position of the phosphoinositide acyl chains, which are found to align with the membrane insertion element. Dual engagement of the FYVE domain with PtdIns(3)P and dodecylphosphocholine micelles yields a 6-fold enhancement of affinity. The additional interaction of phosphatidylserine with a conserved basic site of the protein further amplifies the micelle binding affinity and dramatically alters the angle of insertion. Thus, the FYVE domain is targeted to endosomes through the synergistic action of stereospecific PtdIns(3)P head group ligation, hydrophobic insertion and electrostatic interactions with acidic phospholipids.Cellular processes including signal transduction, vesicular trafficking, and cytoskeletal rearrangement require selective recruitment of proteins to membrane surfaces. Well established mechanisms for localizing cytosolic proteins to membranes include electrostatic interactions through a basic peptide sequence, anchoring by covalently attached acyl chains, and association with the cytoplasmic domains of transmembrane proteins (reviewed in Refs. 1 and 2). (4), PDZ (5), and PTB (6) domains. Although the majority of these domains can interact with several PIs, the FYVE domain is remarkably selective for PtdIns(3)P (7-9). In addition to PI ligation, these domains often insert hydrophobic elements into the membrane bilayer, as has been demonstrated for the C2 (10), ENTH (11), FERM (12), FYVE (13), and PX (14) domains and the vinculin tail (15). Insertion into the membrane can be accompanied by interactions with multiple lipid head groups. For example, the PX domain of the p47 subunit of the NADPH oxidase binds cooperatively to PtdIns(3)P and phosphatidic acid (16), and the vinculin tail co-ligates phosphatidylinositol 4,5-bisphosphate and PtdSer (15). Although it is becoming evident that insertion of proteins into membranes is widespread, the three-dimensional orientations and quantitative binding properties remain challenging to characterize. The most common electron paramagnetic resonance and fluorescence approaches have provided important insights (17-20) but require covalent attachment of paramagnetic groups to various positions of the protein or mutations of residues. The inevitable effects of these modifications on lipi...

show abstract

“…Furthermore, this protein has a well-defined binding surface in comparison with other known PIP-binding domain-containing proteins, indicating that the adamantyl groups are indeed useful replacements of the naturally occurring long-chain fatty acids. [19] …”

Section: Resultsmentioning

confidence: 99%

Synthesis of Highly Water-Soluble Adamantyl Phosphoinositide Derivatives

et al. 2015

View full text Add to dashboard Cite

Phosphatidylinositol phosphates are key regulators of cell signalling pathways and membrane trafficking in eukaryotic cells, and there is a need for new chemical probes to further understand how they interact with lipid-binding proteins. Here, the synthesis of phosphatidylinositol phosphate analogues containing adamantyl carboxylic ester groups, in place of the natural lipid side chains, is described. These derivatives are considerably more soluble in water than analogues containing other lipid side chains and do not form large aggregates such as liposomes or micelles. These adamantyl analogues bind to known phosphoinositide-binding proteins with similar affinities to native ligands and will facilitate future studies on the substrate specificities of these proteins involving cocrystallisation studies with proteins.

show abstract

Crystal Structure of the VHS and FYVE Tandem Domains of Hrs, a Protein Involved in Membrane Trafficking and Signal Transduction

Cited by 166 publications

References 36 publications

Subunit H of the V-ATPase Involved in Endocytosis Shows Homology to β-Adaptins

Subunit H of the V-ATPase Involved in Endocytosis Shows Homology to β-Adaptins

Multivalent Mechanism of Membrane Insertion by the FYVE Domain

Synthesis of Highly Water-Soluble Adamantyl Phosphoinositide Derivatives

Contact Info

Product

Resources

About