Covalent and Noncovalent Intermediates of an NAD Utilizing Enzyme, Human CD38

Liu, Qun; Kriksunov, I.A.; Jiang, Hong; Graeff, Richard; Lin, Hening; Lee, Hon Cheung; Hao, Quan

doi:10.1016/j.chembiol.2008.08.007

Cited by 41 publications

(62 citation statements)

References 52 publications

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“…This observation was also seen in the structures of human CD38, the homolog of Aplysia cyclase. [18][19][20] The results represent the first time ever that a fully active Aplysia cyclase has been observed with an NAD + molecule, its natural substrate, or an NGD + molecule, a close analog of NAD + , bound at the active site. The results also validate our approach of selecting crystals with multiple cyclase molecules in the asymmetric unit for analysis.…”

Section: Overview Of Complex Structuresmentioning

confidence: 87%

“…14 To decipher details of the cyclization reaction by Aplysia cyclase and to understand the structural differences conferring its homolog human CD38 to be more of an NAD + hydrolase than cyclase (i.e., converting NAD + to a linear ADPR instead of the cyclic product, cADPR), we have performed extensive kinetic crystallographic studies to attempt to arrest the reaction at specific steps. 12,13,[15][16][17][18][19][20][21] In most cases, the crystals of both proteins contained two molecules per asymmetric unit, and even though different states of the reaction intermediate can be captured in the two molecules, it could not provide us with a comprehensive picture about the progress of the reaction. Interestingly, Aplysia cyclase and human CD38 have the ability to crystallize in different crystal forms, some of which contain multiple subunits of the protein in the asymmetric unit.…”

Section: Introductionmentioning

confidence: 98%

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Structural Studies of Intermediates along the Cyclization Pathway of Aplysia ADP-Ribosyl Cyclase

Kotaka

Graeff

Chen

et al. 2012

Journal of Molecular Biology

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Section: Overview Of Complex Structuresmentioning

confidence: 87%

Section: Introductionmentioning

confidence: 98%

Structural Studies of Intermediates along the Cyclization Pathway of Aplysia ADP-Ribosyl Cyclase

Kotaka

Graeff

Chen

et al. 2012

Journal of Molecular Biology

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“…To visualize the cyclization process, however, we had to choose an appropriate substrate that can form a stable complex with the cyclase without being consumed and released as product. One possibility is ara-2ЈF-NMN, which has been shown to form a covalent intermediate with CD38 and inhibit its enzyme activity (33,34). However, ara-2ЈF-NMN cannot form a cyclic product because it is missing the adenine group.…”

Section: Resultsmentioning

confidence: 99%

Mechanism of Cyclizing NAD to Cyclic ADP-ribose by ADP-ribosyl Cyclase and CD38

Graeff

Liu

Kriksunov

et al. 2009

Journal of Biological Chemistry

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Mammalian CD38 and its Aplysia homolog, ADP-ribosyl cyclase (cyclase), are two prominent enzymes that catalyze the synthesis and hydrolysis of cyclic ADP-ribose (cADPR), a Ca 2؉ messenger molecule responsible for regulating a wide range of cellular functions. Although both use NAD as a substrate, the cyclase produces cADPR, whereas CD38 produces mainly ADPribose (ADPR). To elucidate the catalytic differences and the mechanism of cyclizing NAD, the crystal structure of a stable complex of the cyclase with an NAD analog, ribosyl-2F-2de-oxynicotinamide adenine dinucleotide (ribo-2-F-NAD), was determined. The results show that the analog was a substrate of the cyclase and that during the reaction, the nicotinamide group was released and a stable intermediate was formed. The terminal ribosyl unit at one end of the intermediate formed a close linkage with the catalytic residue (Glu-179), whereas the adenine ring at the other end stacked closely with Phe-174, suggesting that the latter residue is likely to be responsible for folding the linear substrate so that the two ends can be cyclized. Mutating Phe-174 indeed reduced cADPR production but enhanced ADPR production, converting the cyclase to be more CD38-like. Changing the equivalent residue in CD38, Thr-221 to Phe, correspondingly enhanced cADPR production, and the double mutation, Thr-221 to Phe and Glu-146 to Ala, effectively converted CD38 to a cyclase. This study provides the first detailed evidence of the cyclization process and demonstrates the feasibility of engineering the reactivity of the enzymes by mutation, setting the stage for the development of tools to manipulate cADPR metabolism in vivo.Cyclic ADP-ribose is a novel cyclic nucleotide with Ca 2ϩ -mobilizing activity targeting the endoplasmic reticulum. Its activity was first described in sea urchin eggs (1, 2), and cADPR 3 has since been established as a second messenger molecule responsible for regulating a wide range of physiological functions, from fission in the dinoflagellate (3) to social behavior in mice (Ref. 4 and reviewed in Refs. 5 and 6). The Aplysia ADPribosyl cyclase (cyclase) was the first protein identified that uses NAD, a linear substrate, and ligates its two ends to produce cADPR, with the release of the terminal moiety, nicotinamide (7). The cyclase is a soluble protein of 30 kDa and is present in large amounts in Aplysia ovotestis (7). It is also present in the neurons of the Aplysia buccal ganglion, where it is responsible for the synthesis of endogenous cADPR and the regulation of the evoked synaptic transmission (8). Recently, it is shown that depolarization of Aplysia neurons induces the translocation of the cyclase from the cytosol into the nucleus, providing a mechanism for fine tuning of nuclear Ca 2ϩ signals in neurons (9). CD38 is the major mammalian homolog of the cyclase and is responsible for regulating a wide range of physiological functions. Deletion of the CD38 gene in mice produces multiple defects, including impairment of insulin secretion (10), neutrophil chemota...

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“…An intermediate is formed. Both covalent [124] and non-covalent [123,124] intermediates of CD38 have been observed, depending on the substrate. With arabinosyl-2′-fluoro-deoxy-nicotinamide adenine dinucleotide (ara-2′F-NAD) as substrate, an analog of NAD, Glu226 forms a covalent linkage with the anomeric carbon (C1R) of the terminal ribose [125], as shown in Figures 3 and 4.…”

Section: Structure and Catalytic Mechanism Of Cd38mentioning

confidence: 99%

Cyclic ADP-ribose and NAADP: fraternal twin messengers for calcium signaling

Lee

2011

Sci. China Life Sci.

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The concept advanced by Berridge and colleagues that intracellular Ca 2+ -stores can be mobilized in an agonist-dependent and messenger (IP 3 )-mediated manner has put Ca 2+ -mobilization at the center stage of signal transduction mechanisms. During the late 1980s, we showed that Ca 2+ -stores can be mobilized by two other messengers unrelated to inositol trisphosphate (IP 3 ) and identified them as cyclic ADP-ribose (cADPR), a novel cyclic nucleotide from NAD, and nicotinic acid adenine dinucleotide phosphate (NAADP), a linear metabolite of NADP. Their messenger functions have now been documented in a wide range of systems spanning three biological kingdoms. Accumulated evidence indicates that the target of cADPR is the ryanodine receptor in the sarco/endoplasmic reticulum, while that of NAADP is the two pore channel in endolysosomes.As cADPR and NAADP are structurally and functionally distinct, it is remarkable that they are synthesized by the same enzyme. They are thus fraternal twin messengers. We first identified the Aplysia ADP-ribosyl cyclase as one such enzyme and, through homology, found its mammalian homolog, CD38. Gene knockout in mice confirms the important roles of CD38 in diverse physiological functions from insulin secretion, susceptibility to bacterial infection, to social behavior of mice through modulating neuronal oxytocin secretion. We have elucidated the catalytic mechanisms of the Aplysia cyclase and CD38 to atomic resolution by crystallography and site-directed mutagenesis. This article gives a historical account of the cADPR/NAADP/CD38-signaling pathway and describes current efforts in elucidating the structure and function of its components.cyclic ADP-ribose, cADPR, NAADP, nicotinic acid adenine dinucleotide phosphate, CD38, ADP-ribosyl cyclase, Calcium mobilization and signaling Citation:Lee H C. Cyclic ADP-ribose and NAADP: fraternal twin messengers for calcium signaling. Sci China Life Sci, 2011Sci, , 54: 699 -711, doi: 10.1007 In 1983, Berridge and colleagues published a study showing that agonists, such as carbachol, can activate Ca 2+ release from non-mitochondrial Ca 2+ stores and the effect is mediated by a Ca 2+ messenger, inositol 1,4,5-trisphosphate (IP 3 ) [1]. The targeted Ca 2+ stores were later identified as the endoplasmic reticulum (ER), which has since been shown to be the major intracellular Ca 2+ stores. This seminal study ushered in the current field of Ca 2+ signaling as we know it and has made the field center stage. IP 3 is derived from a phospholipid, phosphatidyl inositol 4,5-bisphosphate, present on the cytoplasmic side of the plasma membrane. Agonist binding to its specific receptor leads to activation of phospholipase C and the hydrolysis of the phospholipid, releasing the head group that is IP 3 . The structure of IP 3 is shown in Figure 1. It has three phosphates on the inositol ring. Both the number of phosphates and the position of the phosphates are critical to the binding of IP 3 to its receptor in the ER. The space-filling model of IP 3 shown in Figu...

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Covalent and Noncovalent Intermediates of an NAD Utilizing Enzyme, Human CD38

Cited by 41 publications

References 52 publications

Structural Studies of Intermediates along the Cyclization Pathway of Aplysia ADP-Ribosyl Cyclase

Structural Studies of Intermediates along the Cyclization Pathway of Aplysia ADP-Ribosyl Cyclase

Mechanism of Cyclizing NAD to Cyclic ADP-ribose by ADP-ribosyl Cyclase and CD38

Cyclic ADP-ribose and NAADP: fraternal twin messengers for calcium signaling

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