45 research outputs found
Profilin 1 is essential for retention and metabolism of mouse hematopoietic stem cells in bone marrow
How stem cells interact with the microenvironment to regulate their cell fates and metabolism is largely unknown. Here we demonstrated that the deletion of the cytoskeleton-modulating protein profilin 1 (pfn1) in hematopoietic stem cell (HSCs) led to bone marrow failure, loss of quiescence, and mobilization and apoptosis of HSCs in vivo. A switch from glycolysis to mitochondrial respiration with increased reactive oxygen species (ROS) level was also observed in HSCs on pfn1 deletion. Importantly, treatment of pfn1-deficient mice with the antioxidant N-acetyl-l-cysteine reversed the ROS level and loss of quiescence of HSCs, suggesting that the metabolism is mechanistically linked to the cell cycle quiescence of stem cells. The actin-binding and proline-binding activities of pfn1 are required for its function in HSCs. Our study provided evidence that pfn1 at least partially acts through the axis of pfn1/Gα13/EGR1 to regulate stem cell retention and metabolism in the bone marrow
Clinical proteomic biomarkers: relevant issues on study design & technical considerations in biomarker development
Proteomic advances in salivary diagnostics
Saliva is identified as functional equivalent to serum, reflecting the physiological state of the body, as well as hormonal, emotional, nutritional and metabolic alterations. The application of mass spectrometry based approaches has allowed a thorough characterization of the saliva proteome and led to the discovery of putative biomarkers. Several salivary biomarkers have been recently explored as potentially useful screening tools in patients diagnosed with metabolic disorders. In this review, we provide an overview of saliva proteomics studies, with a focus on diabetes, and we explore the evidence for the utility of well identified markers for the diagnosis and monitoring of the disease. Emerging approaches in salivary diagnostics that may significantly advance the field of diabetes research are also highlighted. © 202
Modular Structure, Local Flexibility and Cold-Activity of a Novel Chitobiase from a Psychrophilic Antarctic Bacterium
The gene archb encoding for the cell-bound chitobiase from the Antarctic Gram-positive bacterium Arthrobacter sp. TAD20 was cloned and expressed in Escherichia coli in a soluble form. The mature chitobiase ArChb possesses four functionally independent domains: a catalytic domain stabilized by Ca(2+), a galactose-binding domain and an immunoglobulin-like domain followed by a cell-wall anchorage signal, typical of cell-surface proteins from Gram-positive bacteria. Binding of saccharides was analyzed by differential scanning calorimetry, allowing to distinguish unequivocally the catalytic domain from the galactose-binding domain and to study binding specificities. The results suggest that ArChb could play a role in bacterium attachment to natural hosts. Kinetic parameters of ArChb demonstrate perfect adaptation to catalysis at low temperatures, as shown by a low activation energy associated with unusually low K(m) and high k(cat) values. Thermodependence of these parameters indicates that discrete amino acid substitutions in the catalytic center have optimized the thermodynamic properties of weak interactions involved in substrate binding at low temperatures. Microcalorimetry also reveals that heat-lability, a general trait of psychrophilic enzymes, only affects the active site domain of ArChb
Modular structure, local flexibility and cold-activity of a novel chitobiase from a psychrophilic Antarctic bacterium
The gene archb encoding for the cell-bound chitobiase from the Antarctic
Gram-positive bacterium Arthrobacter sp. TAD20 was cloned and expressed
in Escherichia coli in a soluble form. The mature chitobiase ArChb
possesses four functionally independent domains. a catalytic domain
stabilized by Ca2+, a,galactose-binding domain and an
immunoglobulin-like domain followed by a cell-wall anchorage signal,
typical of cell-surface proteins from Gram-positive bacteria. Binding of
saccharides was analyzed by differential scanning calorimetry, allowing
to distinguish unequivocally the catalytic domain from the
galactose-binding domain and to study binding specificities. The results
su,,est that ArChb could play a role in bacterium attachment to natural
hosts. Kinetic parameters of ArChb demonstrate perfect adaptation to
catalysis at low temperatures, as shown by a low activation energy
associated with unusually low K-m and high k(cat) values.
Thermodependence of these parameters indicates that discrete amino acid
substitutions in the catalytic center have optimized the thermodynamic
properties of weak interactions involved in substrate binding at low
temperatures. Microcalorimetry also reveals that heat-lability, a
general trait of psychrophilic enzymes, only affects the active site
domain of ArChb. (C) 2001 Academic Press