MARCKS-related Protein (MRP) Is a Substrate for the Leishmania major Surface Protease Leishmanolysin (gp63)

Myristoylated alanine-rich C kinase substrate (MARCKS) and MARCKS-related protein (MRP; MacMARCKS) are protein kinase C substrates in diverse cell types. Activation of murine macrophages by cytokines increases MRP expression, but infection with Leishmania promastigotes during activation results in M...

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Bibliographic Details
Published in:The Journal of biological chemistry 1999-09, Vol.274 (36), p.25411-25418
Main Authors: Corradin, Sally, Ransijn, Adriana, Corradin, Giampietro, Roggero, Mario A., Schmitz, Arndt A.P., Schneider, Pascal, Mauël, Jacques, Vergères, Guy
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Hydrolysis
Leishmania major
Leishmania major - enzymology
Mass Spectrometry
Membrane Proteins - metabolism
Metalloendopeptidases - chemistry
Metalloendopeptidases - metabolism
Molecular Sequence Data
Protozoan Proteins - metabolism
Substrate Specificity
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Summary:Myristoylated alanine-rich C kinase substrate (MARCKS) and MARCKS-related protein (MRP; MacMARCKS) are protein kinase C substrates in diverse cell types. Activation of murine macrophages by cytokines increases MRP expression, but infection with Leishmania promastigotes during activation results in MRP depletion. We therefore examined the effect of Leishmania major LV39 on recombinant MRP. Both live promastigotes and a soluble fraction of LV39 lysates degraded MRP to yield lower molecular weight fragments. Degradation was independent of MRP myristoylation and was inhibited by protein kinase C-dependent phosphorylation of MRP. MRP was similarly degraded by purified leishmanolysin (gp63), a Leishmania surface metalloprotease. Degradation was evident at low enzyme/substrate ratios, over a broad pH range, and was inhibited by 1,10-phenanthroline and by a hydroxamate dipeptide inhibitor of leishmanolysin. Using mass spectrometric analysis, cleavage was shown to occur within the effector domain of MRP between Ser92 and Phe93, in accordance with the substrate specificity of leishmanolysin. Moreover, an MRP construct in which the effector domain had been deleted was resistant to cleavage. Thus, Leishmania infection may result in leishmanolysin-dependent hydrolysis of MRP, a major protein kinase C substrate in macrophages.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.274.36.25411