2,480 research outputs found
Intervista a Dario Maccarrone
No full textIl contributo riporta gli esiti dell'intervista effettuata a Dario Maccarrone, curata dall'autore dell'articolo, in riferimento al ruolo delle startup nei territori interni
MODULATION OF THE ENZYMATIC ACTIVITY AND MEMBRANE BINDING PROPERTIES OF SOYBEAN LIPOXYGENASE-1 THROUGH LIMITED PROTEOLYSIS AND METAL SUBSTITUTION
No full textLipoxygenases are non-heme, non-sulfur iron containing enzymes that catalyze the dioxygenation of polyunsatured fatty acids containing one or more pentadiene systems to the corresponding hydroperoxy derivatives. Structural studies in solution of the mammalian and plant enzyme revealed that the latter has a more stable and compact conformation1. As yet, metal atom extraction, reconstitution and substitution with vicariate metals have not been successfully applied to soybean lipoxygenase-1, because of the highly buried position of the iron atom within the active site. Tryptic digestion of lipoxygenase-1 and the subsequent isolation of the 60 kDa C-terminal region allowed to generate a “mini-lipoxygenase-1 (miniLOX)” that retains the catalytically active iron, but in a more accessible position2. In this study, we investigated by near-UV-circular dichroism and fluorescence spectroscopies the structural and functional effects of iron removal, reconstitution and vicariation in miniLOX. Moreover, we report the kinetic analysis and the membrane binding ability of the apo- and metal-substituted forms of miniLOX, using fluorescence resonance energy transfer and monolamellar vesicles. Taken together, these data demonstrate an unprecedented structural role of iron, which is involved not only in the catalytic activity but also in the membrane binding ability of lipoxygenase-1.1. Dainese E. Sabatucci A. van Zadelhoff G. Angelucci C. B. Vachette P., Veldink G. Finazzi Agrò A. and Maccarrone M. (2005). J. Mol. Biol. 349, 143-152.2. Maccarrone M. Salucci M. L. van Zadelhoff G. Malatesta F. Veldink, G. Vliegenthart J. F. and Finazzi Agrò A. (2001). Biochemistry 40, 6819-6827.[...
Chapter 1. Quest for Magic Bullets to Solve the Endocannabinoid Puzzle
No full textCannabis (Cannabis sativa and Cannabis indica) is one of the oldest cultivated plant. Its extracts contain >110 (phyto)cannabinoids (pCBs), some of which appear of clear therapeutic interest. Among them, ?9-tetrahydrocannabinol (THC) and cannabidiol (CBD) hold promise to treat several human diseases, both within the central nervous system and at the periphery. Yet, the complexity of cannabis makes it challenging to really exploit a specific active ingredient for a specific therapeutic purpose. This complexity of the plant extracts is mirrored, or even exceeded, by the complexity of the molecular targets that pCBs can find in our body, most of which belong to the so-called "endocannabinoid (eCB) system". Here, I describe the main components of this signalling system, in order to appreciate how challenging it is to develop drugs that can hit specifically each of them, with a benefit for human health
Altered anti-inflammatory resolvin E1 in an in vivo model of age-related macular degeneration
No full textDifferential regulation of fatty acid amide hydrolase promoter in human immune cells and neuronal cells by leptin and progesterone
No full textWe have shown recently that in human T lymphocytes, leptin stimulates activity and expression of the endocannabinoid-degrading enzyme fatty acid amide hydrolase (FAAH), through STAT3 (signal transducer and activator of transcription 3) and its CRE (cAMP response element)-like transcriptional target in the FAAH promoter [Maccarrone, M., Di Rienzo, M., Finazzi-Agro, A., & Rossi, A. (2003) J. Biol. Chem. 278, 13318-13324]. We have also shown that progesterone, alone or additively with leptin, up-regulates the FAAH gene in human T-cells, through the Ikaros transcription factor [Maccarrone, M., Bari, M., Di Rienzo, M., Finazzi-Agro, A., & Rossi, A. (2003) J. Biol. Chem. 278, 32726-32732]. Here, we extend these observations to immortalized human lymphoma U937 cells, where stimulation of FAAH by leptin (up to approximate to 300% of the controls) involves binding to a leptin receptor (K-d = 2.0 +/- 0.1 nm, B-max = 382 +/- 5 fmol.mg protein(-1), apparent molecular mass of approximate to 110 kDa), and stimulation by progesterone involves an intracellular receptor of approximate to 120 kDa. Unlike FAAH, the other proteins of the endocannabinoid system are not modulated by the two hormones. Interestingly, human neuroblastoma CHP100 cells also have a leptin receptor (approximate to 110 kDa, K-d = 2.2 +/- 0.2 n<smallcapitals>m</smallcapitals>, B-max = 339 +/- 8 fmol.mg protein(-1)), a progesterone receptor (approximate to 120 kDa), STAT3 and Ikaros, yet their FAAH is not activated by leptin or progesterone. These data, corroborated by transient expression and electrophoretic mobility-shift assays, demonstrate an unprecedented cell-specific regulation of the FAAH gene, which has important implications for the control of tone and activity of AEA along the neuroimmune axis
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