Crystal Structure of β-Arrestin at 1.9 Å

Han, May; Gurevich, Vsevolod V.; Vishnivetskiy, Sergey A.; Sigler, Paul B.; Schubert, Carsten J.

doi:10.1016/s0969-2126(01)00644-x

Cited by 348 publications

(274 citation statements)

References 64 publications

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“…The crystal structure of ␤-arrestin 1 has been determined (44). ␤-Arrestin is formed from two major domains, called the N-and C-domains, each of which is composed of a seven-strand ␤-sandwich.…”

Section: Discussionmentioning

confidence: 99%

“…5). The crystal structure of ␤-arrestin 1 (44) indicates that these two regions do not juxtapose each other. One possibility is that they may provide distinct sites for interaction with the two regions in PDE4D5 shown here to confer interaction with ␤-arrestin.…”

Section: Discussionmentioning

confidence: 99%

“…One possibility is that they may provide distinct sites for interaction with the two regions in PDE4D5 shown here to confer interaction with ␤-arrestin. One of the two regions in ␤-arrestin 2, located within amino acids 1-28, contains two pairs of positively charged amino acids, namely Lys 11 /Lys 12 and Lys 25 /Arg 26 , which are located at the ends of distinct ␤-sheets in ␤-arrestin 1 (44). The mutation of these amino acids to the neutral alanine ablates the ability of ␤-arrestin 2 to interact with PDE4D5 (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Analysis of the ␤-arrestin 1 crystal structure (44) shows it to be composed of two major regions, known as the N-and C-domains, each of which is composed of a seven-strand ␤-sandwich (Fig. 5a).…”

Section: Identification Of Two Regions In ␤-Arrestin That Are Involvementioning

confidence: 99%

“…5a). These regions are considered to have distinct functional attributes (44). The N-domain binds to appropriate phosphorylated GPCRs and SRC, whereas the C-domain is involved in binding to JNK3, clathrin, and the ␤ 2 -adaptin subunit of the AP2 complex.…”

Section: Identification Of Two Regions In ␤-Arrestin That Are Involvementioning

confidence: 99%

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The Unique Amino-terminal Region of the PDE4D5 cAMP Phosphodiesterase Isoform Confers Preferential Interaction with β-Arrestins

Bolger

McCahill

Huston

et al. 2003

Journal of Biological Chemistry

101

139

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Isoproterenol challenge of Hek-B2 cells causes a transient recruitment of the endogenous PDE4D isoforms found in these cells, namely PDE4D3 and PDE4D5, to the membrane fraction. PDE4D5 provides around 80% of the total PDE4D protein so recruited, although it only comprises about 40% of the total PDE4D protein in Hek-B2 cells. PDE4D5 provides about 80% of the total PDE4D protein found associated with ␤-arrestins immunopurified from Hek-B2, COS1, and A549 cells as well as cardiac myocytes, whereas its overall level in these cells is between 15 and 50% of the total PDE4D protein. Truncation analyses indicate that two sites in PDE4D5 are involved in mediating its interaction with ␤-arrestins, one associated with the common PDE4 catalytic region and the other located within its unique amino-terminal region. Truncation analyses indicate that two sites in ␤-arrestin 2 are involved in mediating its interaction with PDE4D5, one associated with its extreme amino-terminal region and the other located within the carboxylterminal domain of the protein. We suggest that the unique amino-terminal region of PDE4D5 allows it to preferentially interact with ␤-arrestins. This specificity appears likely to account for the preferential recruitment of PDE4D5, compared with PDE4D3, to membranes of Hek-B2 cells and cardiac myocytes upon challenge with isoproterenol.The heptahelical ␤-adrenergic receptors (␤ 2 ARs) 1 act by coupling through the G-protein G s to stimulate adenylyl cyclase and thereby increase intracellular cAMP concentrations (1-4). It is now well established that the rapid uncoupling of this response is achieved by the action of the G-protein receptorcoupled kinase 2 (2). This phosphorylates the carboxyl-terminal tail of the plasma membrane-associated ␤ 2 AR, allowing the recruitment of ␤-arrestins from the cytosol (5-7). It is this recruitment of ␤-arrestin, rather than phosphorylation per se, that elicits uncoupling, presumably by sterically blocking coupling of the ␤ 2 AR to G s . The importance of ␤-arrestin interaction to G-protein-coupled receptors (GPCRs) in vivo has been clearly established in knockout mouse models (8 -10).The attenuation of cAMP signaling is also achieved through the action of phosphodiesterases (PDEs) that are able to hydrolyze cAMP to 5Ј-AMP (11-17). Since they provide the sole route for degradation of cAMP in cells, they are poised to play a key role in controlling cAMP signaling. Multiple genes encode a large superfamily of PDEs, which differ in their regulatory and kinetic properties. Of these, the PDE4 cAMP-specific phosphodiesterase family (13-16) has recently attracted much interest, since PDE4-selective inhibitors are currently being developed as potential therapeutic agents for various inflammatory diseases of the respiratory system, such as asthma and chronic obstructive pulmonary disease (18 -21). The PDE4 enzyme family is encoded by four genes (PDE4A, -B, -C, and -D), which generate over 16 different isoforms through the use of distinct promoters and alternative mRNA splicing (12,13,...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Identification Of Two Regions In ␤-Arrestin That Are Involvementioning

confidence: 99%

Section: Identification Of Two Regions In ␤-Arrestin That Are Involvementioning

confidence: 99%

See 3 more Smart Citations

The Unique Amino-terminal Region of the PDE4D5 cAMP Phosphodiesterase Isoform Confers Preferential Interaction with β-Arrestins

Bolger

McCahill

Huston

et al. 2003

Journal of Biological Chemistry

101

139

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Arrestin Expression in E. coli and Purification

et al. 2014

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Purified arrestin proteins are necessary for biochemical, biophysical, and crystallographic studies of these versatile regulators of cell signaling. Here we describe a basic protocol for expression in E. coli and purification of tag-free wild type and mutant arrestins. The method includes ammonium sulfate precipitation of arrestins from cell lysates, followed by heparin-Sepharose chromatography. Depending on the arrestin type and/or mutations, this step is followed by Q-Sepharose or SP-Sepharose chromatography. In many cases the non-binding column is used as a pre-filter to bind contaminants without retaining arrestin. In some cases both chromatographic steps need to be performed sequentially to achieve high purity. Purified arrestins can be concentrated up to 10 mg/ml, remain fully functional, and can withstand several cycles of freezing and thawing, provided that overall salt concentration is kept at or above physiological levels.

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Eine kurze Geschichte der G‐Protein‐gekoppelten Rezeptoren (Nobel‐Aufsatz)

Lefkowitz

2013

Angewandte Chemie

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Crystal Structure of β-Arrestin at 1.9 Å

Cited by 348 publications

References 64 publications

The Unique Amino-terminal Region of the PDE4D5 cAMP Phosphodiesterase Isoform Confers Preferential Interaction with β-Arrestins

The Unique Amino-terminal Region of the PDE4D5 cAMP Phosphodiesterase Isoform Confers Preferential Interaction with β-Arrestins

Arrestin Expression in E. coli and Purification

Eine kurze Geschichte der G‐Protein‐gekoppelten Rezeptoren (Nobel‐Aufsatz)

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