Lenz-Majewski syndrome (LMS) is a syndrome of intellectual disability and multiple congenital anomalies that features generalized craniotubular hyperostosis. By using whole-exome sequencing and selecting variants consistent with the predicted dominant de novo etiology of LMS, we identified causative heterozygous missense mutations in PTDSS1, which encodes phosphatidylserine synthase 1 (PSS1). PSS1 is one of two enzymes involved in the production of phosphatidylserine. Phosphatidylserine synthesis was increased in intact fibroblasts from affected individuals, and end-product inhibition of PSS1 by phosphatidylserine was markedly reduced. Therefore, these mutations cause a gain-of-function effect associated with regulatory dysfunction of PSS1. We have identified LMS as the first human disease, to our knowledge, caused by disrupted phosphatidylserine metabolism. Our results point to an unexplored link between phosphatidylserine synthesis and bone metabolism.

• MeSH
• dánio pruhované embryologie   genetika   MeSH
• dítě MeSH
• embryo nesavčí MeSH
• fibroblasty metabolismus   MeSH
• fosfatidylseriny biosyntéza   genetika   MeSH
• hyperostóza MeSH
• kultivované buňky MeSH
• lidé MeSH
• mladiství MeSH
• mnohočetné abnormality genetika   MeSH
• molekulární sekvence - údaje MeSH
• mutace * MeSH
• nanismus MeSH
• syndrom MeSH
• transferasy dusíkatých skupin genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• fosfatidylseriny MeSH
• phospholipid serine base exchange enzyme MeSH Prohlížeč
• transferasy dusíkatých skupin MeSH

The frequency of Candida infections has increased in recent years and it has been accompanied by a significant rise in morbidity and mortality. The secretion of aspartic proteases by Candida spp. was demonstrated to be one of the virulence determinants. Candida albicans is classified as the major human pathogen in the genus Candida. However, other species of this genus have been found to cause an increasing number of candidiases. We isolated secreted aspartic proteases (Saps) of C. albicans (Sap2p), C. tropicalis (Sapt1p), C. parapsilosis (Sapp1p), and C. lusitaniae (Saplp) from culture media. All the isolated proteases were N-terminally sequenced. Their specific proteolytic activities and sensitivity to series of peptidomimetic inhibitors modified in the type of scissile bond replacement as well as in the N- and C-termini were analyzed. The most divergent substrate specificity was observed for the Sap of C. tropicalis. The specificity of Sap of C. lusitaniae is most closely related to that of Sap of C. parapsilosis. We designed and prepared an inhibitor containing phenylstatine isoster that was equipotent towards all four proteases within the range of 10-10-10-9 M. The HIV-1 protease inhibitors ritonavir, saquinavir, indinavir, and nelfinavir were also tested for the inhibition of four Saps. Only ritonavir and saquinavir inhibited Sap2p, Sapt1p, Sapp1p, and Saplp in micromolar concentrations.

• MeSH
• aspartátové endopeptidasy antagonisté a inhibitory   genetika   izolace a purifikace   metabolismus   MeSH
• Candida albicans enzymologie   MeSH
• Candida enzymologie   MeSH
• inhibitory proteas chemie   farmakologie   MeSH
• kinetika MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• pepstatiny chemie   farmakologie   MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie aminokyselin MeSH
• substrátová specifita MeSH
• techniky in vitro MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• aspartátové endopeptidasy MeSH
• inhibitory proteas MeSH
• pepstatin MeSH Prohlížeč
• pepstatiny MeSH
• Streptomyces pepsin inhibitor MeSH Prohlížeč

This study was designed to analyze the association of Nramp1 and/or Lps genes with differential protein expression in macrophages in order to select candidate proteins that might be related to resistance/susceptibility to various microbial infections under the control of the Nramp1 and/or Lps genes. The macrophage cell lines derived from bone marrow of Nramp1 or Lps congenic mice were utilized and high-resolution two-dimensional electrophoreis (2-DE), separating proteins according to their charge and size, was used as a window into alterations in gene expression and a means to compare the macrophages carrying a resistant allele of Nramp1 gene and/or normal allele of Lps gene, with their counterparts carrying either a susceptible allele of Nramp1 or defective allele of the Lps gene. We demonstrate that the changes of constitutive levels of two proteins named according to their isoelectric point/molecular weight (pI/Mr), p6.6/25 and p7.0/22, discriminate satisfactorily not only the macrophages congenic at the Nramp1 gene but also the macrophages congenic at the Lps gene, thus reflecting their common genotype (Nramp1r, Lps[n]). Furthermore, the decreased constitutive levels of these proteins in macrophages carrying a defective allele of Lps but preserving a resistant allele of Nramp1 can be, at least in part, restored by stimulation with interferon gamma or lipopolysaccharide. 2-DE immunoblot analysis identified the p7.0/22 protein as manganese superoxide dismutase. Bcl-2 appears to be the best candidate for p6.6/25 as suggested by 2-DE quantitative alterations and Western blot analysis. These proteins are important in the regulation of intracellular redox balance and the regulation of apoptosis in macrophages and their alterations might reflect closely the transport functions of ions or other charged substrates suggested for Nramp1 protein.

• MeSH
• buněčné linie MeSH
• křížení genetické MeSH
• lipopolysacharidy * MeSH
• makrofágy chemie   cytologie   MeSH
• membránové proteiny genetika   MeSH
• molekulární sekvence - údaje MeSH
• myši MeSH
• přirozená imunita genetika   MeSH
• proteiny přenášející kationty * MeSH
• proteiny analýza   genetika   MeSH
• sekvence aminokyselin MeSH
• transportní proteiny genetika   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• lipopolysacharidy * MeSH
• membránové proteiny MeSH
• natural resistance-associated macrophage protein 1 MeSH Prohlížeč
• proteiny přenášející kationty * MeSH
• proteiny MeSH
• transportní proteiny MeSH