A Novel Mechanism for Adenylyl Cyclase Inhibition from the Crystal Structure of Its Complex with Catechol Estrogen

Steegborn, Clemens; Litvin, Tatiana N.; Hess, Kenneth C.; Capper, Austin B.; Taussig, Ronald; Buck, Jochen; Levin, Lonny R.; Wu, Hao

doi:10.1074/jbc.m507144200

Cited by 68 publications

(98 citation statements)

References 42 publications

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“…Based on these results, we proposed a modified model for the catalytic role of the α1 and β7/8 movements: substrate binding fully opens the active site, and the subsequent closure drives catalysis by separating the products cAMP and pyrophosphate. 42 Consistent with this model, the tmAC-G s α complexes with P-site inhibitors, which presumably resemble an enzyme/product complex, are similar to the closed enzyme conformation. 63 During active site closure, Pγ and Pβ would be shifted toward the active site exit.…”

Section: The Catalytic Mechanism Of Class III Cyclasessupporting

confidence: 54%

“…Additionally, mammalian sAC and the CyaC cyclase from the cyanobacteria Spirulina platensis are similarly regulated; both are synergistically activated by bicarbonate and calcium ions. 20,42 Bicarbonate-regulated sAC-like cyclases are also found in Anabaena PCC 7120, Stigmatella aurantiaca, Mycobacterium tuberculosis, 43 and the thermophilic bacterium Chloroflexus aurantiacus, 44 and sAC-like proteins were cloned and expressed from sea urchin 45 and Plasmodium falciparum, the parasite responsible for the most lethal form of malaria. 46 Database searches identify sAC-like ACs in the genomes of all mammals studied (human, chimpanzee, dog, cow, rabbit, mouse, and rat), as well as in chicken, ciona intestinalis, and insects (mosquito and honey bee).…”

Section: Evolution Of Mammalian Adenylyl Cyclasesmentioning

confidence: 99%

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Molecular Details of cAMP Generation in Mammalian Cells: A Tale of Two Systems

Kamenetsky

Middelhaufe

Bank

et al. 2006

Journal of Molecular Biology

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295

340

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The second messenger cAMP has been extensively studied for half a century, but the plethora of regulatory mechanisms controlling cAMP synthesis in mammalian cells is just beginning to be revealed. In mammalian cells, cAMP is produced by two evolutionary related families of adenylyl cyclases, soluble adenylyl cyclases (sAC) and transmembrane adenylyl cyclases (tmAC). These two enzyme families serve distinct physiological functions. They share a conserved overall architecture in their catalytic domains and a common catalytic mechanism, but they differ in their sub-cellular localizations and responses to various regulators. The major regulators of tmACs are heterotrimeric G proteins, which transduce extracellular signals via G protein-coupled receptors. sAC enzymes, in contrast, are regulated by the intracellular signaling molecules bicarbonate and calcium. Here, we discuss and compare the biochemical, structural and regulatory characteristics of the two mammalian AC families. This comparison reveals the mechanisms underlying their different properties but also illustrates many unifying themes for these evolutionary related signaling enzymes.

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Section: The Catalytic Mechanism Of Class III Cyclasessupporting

confidence: 54%

Section: Evolution Of Mammalian Adenylyl Cyclasesmentioning

confidence: 99%

Molecular Details of cAMP Generation in Mammalian Cells: A Tale of Two Systems

Kamenetsky

Middelhaufe

Bank

et al. 2006

Journal of Molecular Biology

Self Cite

295

340

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“…OaPAC activity (∼9 pmol·min −1· μg −1 protein) was as high as that of PAC from Euglena measured under the same conditions (18). Adenylate cyclases require a catalytic metal ion coordinated by an aspartate residue (19,20), and sequence comparisons show the equivalent residue in OaPAC is Asp-200. Replacing this residue with asparagine abolishes any activity under all conditions (Fig.…”

Section: Resultsmentioning

confidence: 92%

Structural insight into photoactivation of an adenylate cyclase from a photosynthetic cyanobacterium

Ohki

Sugiyama

Kawai

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

106

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Cyclic-AMP is one of the most important second messengers, regulating many crucial cellular events in both prokaryotes and eukaryotes, and precise spatial and temporal control of cAMP levels by light shows great promise as a simple means of manipulating and studying numerous cell pathways and processes. The photoactivated adenylate cyclase (PAC) from the photosynthetic cyanobacterium Oscillatoria acuminata (OaPAC) is a small homodimer eminently suitable for this task, requiring only a simple flavin chromophore within a blue light using flavin (BLUF) domain. These domains, one of the most studied types of biological photoreceptor, respond to blue light and either regulate the activity of an attached enzyme domain or change its affinity for a repressor protein. BLUF domains were discovered through studies of photo-induced movements of Euglena gracilis, a unicellular flagellate, and gene expression in the purple bacterium Rhodobacter sphaeroides, but the precise details of light activation remain unknown. Here, we describe crystal structures and the light regulation mechanism of the previously undescribed OaPAC, showing a central coiled coil transmits changes from the light-sensing domains to the active sites with minimal structural rearrangement. Site-directed mutants show residues essential for signal transduction over 45 Å across the protein. The use of the protein in living human cells is demonstrated with cAMP-dependent luciferase, showing a rapid and stable response to light over many hours and activation cycles. The structures determined in this study will assist future efforts to create artificial light-regulated control modules as part of a general optogenetic toolkit.optogenetics | X-ray crystallography | blue light | allostery

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“…In addition, Fig. 2D shows that 2-hydroxyestradiol, a selective sAC inhibitor [28] used at 150 µM, the highest concentration that does not induce by itself the sperm AR (results not shown), partially inhibits the jelly induced AR (~30%) as well as the increase in cAMP levels (~42%). The inactive analog 17-β estradiol served as control.…”

Section: Localization Of Susacmentioning

confidence: 99%

Particulate and soluble adenylyl cyclases participate in the sperm acrosome reaction

Beltrán

Vacquier

Moy

et al. 2007

Biochemical and Biophysical Research Communications

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AbstractcAMP is important in sea urchin sperm signaling, yet the molecular nature of the adenylyl cyclases (ACs) involved remained unknown. These cells were recently shown to contain an ortholog of the mammalian soluble adenylyl cyclase (sAC). Here, we show that sAC is present in the sperm head and as in mammals is stimulated by bicarbonate. The acrosome reaction (AR), a process essential for fertilization, is influenced by the bicarbonate concentration in seawater. By using functional assays and immunofluorescence techniques we document that sea urchin sperm also express orthologs of multiple isoforms of transmembrane ACs (tmACs). Our findings employing selective inhibitors for each class of AC indicate that both sAC and tmACs participate in the sperm acrosome reaction. KeywordsAcrosome reaction; cAMP; Adenylyl cyclases; Sea urchin; Soluble adenylyl cyclase; Sperm; fertilization cAMP, an important second messenger in cell signaling, is fundamental in sperm physiology. Sperm must swim, find the egg and undergo the acrosome reaction (AR) to fertilize it [1]. cAMP levels influence sperm ionic fluxes, motility and the AR [2].Two types of adenylyl cyclases (ACs) are expressed in mammals, transmembrane ACs (tmACs) and soluble AC (sAC) molecularly identified in rat testis [3] and sperm [4]. Restricted to the plasma membrane, tmACs are regulated by heterotrimeric G-proteins and stimulated by forskolin [5]. In contrast, sAC is stimulated by bicarbonate [6] and Ca 2+ [7,8]. In addition to sAC, mouse [9] and human [10] sperm apparently also contain tmACs.Why AC activity, cAMP levels, and cyclic AMP-dependent protein kinase (PKA) activity increase [11] Sea urchin sperm were believed to have only one type of AC and no G proteins [11,13].Recently an ortholog of the mammalian sperm sAC was cloned and sequenced from a sea urchin Strongylocentrotus purpuratus testis cDNA library (susAC). Its activity is higher in the presence of Mn 2+ than Mg 2+ , and is stimulated by pH [18]. Furthermore, sea urchin sperm posses G-proteins [15,19,20]. Here, we show by using specific activators and inhibitors for transmembrane and soluble ACs that both ACs participate in the sperm AR. Moreover, we document the presence of several tmACs and their differential distribution in sperm by immunolocalization and Western blot experiments. PCRs were performed using testis cDNA as template and two oligonucleotide primers. A degenerate sense 5′-TTYGYIGAYATHWSIGG NTT-3′based on rat sAC (Rattus norvegicus), NP067716.1 and an anti-sense 5′-ACGGCTGGCTACATTGACAG-3′; exact match from S. purpuratus coelomocytes AC R61912 EST 030. A 630-bp PCR product was obtained corresponding to one of the two copies of the mammalian adenylyl/guanylyl cyclase catalytic domain. Screening an S. purpuratus testis cDNA Lambda Zap library (Stratagene) with this product yielded two clones containing 26% of the SpAC2 (XP_780688; 479-677) and 25% of the SpAC9 (XP_798394; 509-690) predicted sequences. In addition, we found the predicted sequences for AC1 (SpAC1, XP_787811), AC3 (SpAC3,...

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A Novel Mechanism for Adenylyl Cyclase Inhibition from the Crystal Structure of Its Complex with Catechol Estrogen

Cited by 68 publications

References 42 publications

Molecular Details of cAMP Generation in Mammalian Cells: A Tale of Two Systems

Molecular Details of cAMP Generation in Mammalian Cells: A Tale of Two Systems

Structural insight into photoactivation of an adenylate cyclase from a photosynthetic cyanobacterium

Particulate and soluble adenylyl cyclases participate in the sperm acrosome reaction

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