ABSTRACT
Multiple Sulfatase Deficiency (MSD) is a rare autosomal recessive disease in which the activities of all sulfatases are reduced; its estimated prevalence is 1:1.4 million births. The disease is caused by mutations in SUMF1, which encodes an enzyme involved in the post-translational modification of sulfatases. The MSD phenotype is a combination of the clinical features found in diseases resulting from a deficiency of the individual sulfatases; i.e., mucopolysaccharidosis II, IIIA, IIID, IVA and VI, metachromatic leukodystrophy, X-linked ichthyosis, and the X-linked recessive form of chondrodysplasia punctata. We describe herein the first case of a Brazilian patient with MSD. The case was initially diagnosed as having mucopolysaccharidosis (MPS), due to skeletal alterations, coarse facial features, and urinary excretion of dermatan sulfate and heparan sulfate. Later, after a detailed biochemical investigation, the diagnosis of MSD was established. The analysis of the SUMF1 showed the patient was a compound heterozygote for two novel mutations (p.R349G and p.F244S). This case illustrates the challenges in the diagnosis of a disease considered rare, such as MSD. We point out that the availability of therapy for certain MPS disorders necessitates correct disease assignment, and the need to exclude the likelihood of MSD.
Subject(s)
Multiple Sulfatase Deficiency Disease/genetics , Mutation/physiology , Sulfatases/genetics , Brain/diagnostic imaging , Brazil , Child, Preschool , Dermatan Sulfate/urine , Diagnosis, Differential , Heparitin Sulfate/urine , Humans , Intellectual Disability/etiology , Male , Multiple Sulfatase Deficiency Disease/enzymology , Oxidoreductases Acting on Sulfur Group Donors , Tomography, X-Ray ComputedABSTRACT
The laticifer fluid of Calotropis procera is rich in proteins and there is evidence that they are involved in the pharmacological properties of the latex. However, not much is known about how the latex-containing proteins are produced or their functions. In this study, laticifer proteins of C. procera were pooled and examined by 1D and 2D electrophoresis, masses spectrometry (MALDI-TOF) and characterized in respect of proteolytic activity and oxidative enzymes. Soluble laticifer proteins were predominantly composed of basic proteins (PI>6.0) with molecular masses varying between 5 and 95 kDa. Proteins with a molecular mass of approximately 26,000 Da were more evident. Strong anti-oxidative activity of superoxide dismutase (EC 1.15.1.1) (1007.74+/-91.89 Ug(-1)DM) and, to a lesser extent ascorbate peroxidase (EC 1.11.1.1) (0.117(d)+/-0.013 microMol H(2)O(2)g(-1)min(-1)), were detected. However, catalase (EC 1.11.1.6) was absent. The strong proteolytic activities of laticifer proteins from C. procera were shown to be shared by at least four distinct cysteine proteinases (EC 3.4.22.16) that were isolated by gel filtration chromatography. Serine and metaloproteinases were not detected and aspartic proteinase activities were barely visible. Chitinases (EC 3.2.1.14) were also isolated in a chitin column and their activities quantified. The presence of these enzymatic activities in latex from C. procera may confirm their involvement in resistance to phytopathogens and insects, mainly in its leaves where the latex circulates abundantly.
Subject(s)
Calotropis/metabolism , Latex/metabolism , Plant Proteins/metabolism , Ascorbate Peroxidases , Chitin/chemistry , Chitin/metabolism , Chitinases/chemistry , Chitinases/metabolism , Chromatography, Affinity , Cysteine Endopeptidases/chemistry , Cysteine Endopeptidases/metabolism , Electrophoresis, Gel, Two-Dimensional , Electrophoresis, Polyacrylamide Gel , Latex/chemistry , Molecular Weight , Peroxidases/chemistry , Peroxidases/metabolism , Plant Proteins/chemistry , Protons , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Superoxide Dismutase/chemistry , Superoxide Dismutase/metabolism , TemperatureABSTRACT
The embryotoxic activity and differential binding of plant-derived carbohydrate-recognizing proteins on sea urchin (Lytechinus variegatus) embryo cells was investigated. IC50 doses for toxicity on larvae development varied from 0.6 up to 96.3 microg ml(-1) and these effects were largely reversed by previously heating the proteins. Changes in the glycoconjungate status of the cell surface were assessed by time-course binding of the proteins during embryogenesis according to their carbohydrate-binding specificity. Glucose/mannose binding-proteins bound embryo cells at the same stage of development, at a similar stage to the N-acetylglucosamine/N-acetylneuraminic acid binding-protein (WGA) and earlier than galactose specific ones. FITC-conjugates of these proteins confirmed the above results and revealed the presence of specific and differential receptors for them. Inhibition assays using inhibitory glycoproteins significantly diminished the labelled patterns of FITC-conjugates. In conclusion, the assayed proteins exhibited embryotoxicity and their binding requirements were useful for following changes in the pattern of cell surface glycoconjugates on embryo cells of sea urchin. This property could be useful in analyzing other cell types.
