Puppo R

References (3)

Title : Characterization of all the lipolytic activities in pancreatin and comparison with porcine and human pancreatic juices - Salhi_2020_Biochimie_169_106
Author(s) : Salhi A , Amara S , Mansuelle P , Puppo R , Lebrun R , Gontero B , Aloulou A , Carriere F
Ref : Biochimie , 169 :106 , 2020
Abstract : Porcine pancreatic extracts (PPE), also named pancreatin, are commonly used as a global source of pancreatic enzymes for enzyme replacement therapy in patients with exocrine pancreatic insufficiency. They are considered as a good substitute of human pancreatic enzymes and they have become a material of choice for in vitro models of digestion. Nevertheless, while the global PPE contents in lipase, protease and amylase activities are well characterized, little is known about individual enzymes. Here we characterized the lipase, phospholipase, cholesterol esterase and galactolipase activities of PPE and compared them with those of porcine (PPJ) and human (HPJ) pancreatic juices. The phospholipase to lipase activity ratio was similar in PPJ and HPJ, but was 4-fold lower in PPE. The galactolipase and cholesterol esterase activities were found at lower levels in PPJ compared to HPJ, and they were further reduced in PPE. The enzymes known to display these activities in HPJ, pancreatic lipase-related protein 2 (PLRP2) and carboxylester hydrolase/bile salt-stimulated lipase (CEH/BSSL), were identified in PPJ using gel filtration experiments, SDS-PAGE and LC-MS/MS analysis. The galactolipase and cholesterol esterase activities of PPE indicated that PLRP2 and CEH/BSSL are still present at low levels in this enzyme preparation, but they were not detected by mass spectrometry. Besides differences between porcine and human enzymes, the lower levels of phospholipase, galactolipase and cholesterol esterase activities in PPE are probably due to some proteolysis occurring during the production process. In conclusion, PPE do not provide a full substitution of the lipolytic enzymes present in HPJ.
ESTHER : Salhi_2020_Biochimie_169_106
PubMedSearch : Salhi_2020_Biochimie_169_106
PubMedID: 31288050

Title : Cyclipostins and Cyclophostin analogs as promising compounds in the fight against tuberculosis - Nguyen_2017_Sci.Rep_7_11751
Author(s) : Nguyen PC , Delorme V , Benarouche A , Martin BP , Paudel R , Gnawali GR , Madani A , Puppo R , Landry V , Kremer L , Brodin P , Spilling CD , Cavalier JF , Canaan S
Ref : Sci Rep , 7 :11751 , 2017
Abstract : A new class of Cyclophostin and Cyclipostins (CyC) analogs have been investigated against Mycobacterium tuberculosis H37Rv (M. tb) grown either in broth medium or inside macrophages. Our compounds displayed a diversity of action by acting either on extracellular M. tb bacterial growth only, or both intracellularly on infected macrophages as well as extracellularly on bacterial growth with very low toxicity towards host macrophages. Among the eight potential CyCs identified, CyC 17 exhibited the best extracellular antitubercular activity (MIC50 = 500 nM). This compound was selected and further used in a competitive labelling/enrichment assay against the activity-based probe Desthiobiotin-FP in order to identify its putative target(s). This approach, combined with mass spectrometry, identified 23 potential candidates, most of them being serine or cysteine enzymes involved in M. tb lipid metabolism and/or in cell wall biosynthesis. Among them, Ag85A, CaeA and HsaD, have previously been reported as essential for in vitro growth of M. tb and/or survival and persistence in macrophages. Overall, our findings support the assumption that CyC 17 may thus represent a novel class of multi-target inhibitor leading to the arrest of M. tb growth through a cumulative inhibition of a large number of Ser- and Cys-containing enzymes participating in important physiological processes.
ESTHER : Nguyen_2017_Sci.Rep_7_11751
PubMedSearch : Nguyen_2017_Sci.Rep_7_11751
PubMedID: 28924204
Gene_locus related to this paper: mycle-ML2603 , myctu-a85a , myctu-a85c , myctu-Rv1399c , myctu-Rv2970c , myctu-Rv3569c , myctu-yt28

Title : Supported inhibitor for fishing lipases in complex biological media and mass spectrometry identification - Delorme_2014_Biochimie_107 Pt A_124
Author(s) : Delorme V , Raux B , Puppo R , Leclaire J , Cavalier JF , Marc S , Kamarajugadda PK , Buono G , Fotiadu F , Canaan S , Carriere F
Ref : Biochimie , 107 Pt A :124 , 2014
Abstract : A synthetic phosphonate inhibitor designed for lipase inhibition but displaying a broader range of activity was covalently immobilized on a solid support to generate a function-directed tool targeting serine hydrolases. To achieve this goal, straightforward and reliable analytical techniques were developed, allowing the monitoring of the solid support's chemical functionalization, enzyme capture processes and physisorption artifacts. This grafted inhibitor was tested on pure lipases and serine proteases from various origins, and assayed for the selective capture of lipases from several complex biological extracts. The direct identification of captured enzymes by mass spectrometry brought the proof of concept on the efficiency of this supported covalent inhibitor. The features and limitations of this "enzyme-fishing" proteomic tool provide new insight on solid-liquid inhibition process.
ESTHER : Delorme_2014_Biochimie_107 Pt A_124
PubMedSearch : Delorme_2014_Biochimie_107 Pt A_124
PubMedID: 25064360