S, the measurement of overall level of carbonyls of plasma proteins may be used as a surrogate marker of oxidative modification of VWF, as shown in the present study. In conclusion, the emerging scenario shows that the oxidative stress in diabetes involves also VWF and is associated with increased presence of UL-VWF multimers that are involved in the genesis of major cardiovascular events in this clinical setting.AcknowledgmentsRDC thanks prof. V. De Filippis (University of Padua, Italy) for valuable suggestions and scientific discussions. The generous gift of recombinant VWF by Dr. Friedrich Scheiflinger (Baxter Innovations GmbH, Vienna, Austria) is gratefully acknowledged.Author ContributionsEnrolled patients and analyzed the data: FZ. Enrolled patients: FM. Analyzed the data and revised the manuscript: GG. Conceived and designed the experiments: RDC SL DP. Performed the experiments: LO SL RDC PR. Analyzed the data: RDC SL LO DP. Contributed reagents/ materials/analysis tools: RDC SL LO. Wrote the paper: RDC.
Multidomain proteins due to their structural complexity require different levels of regulatory mechanisms for executing cellular functions efficiently within a specified time period. Allosteric modulation of conformations is one such mechanism which often helps a protein to regulate a order Tubastatin-A functional behaviour such as for an enzyme to attain an active functional state upon ligand or substrate binding. In allostery, sometimes there are large conformational changes that require significant rotations and translations of individual domains at the timescales of microsecond to millisecond. While in some other cases, minimal structural perturbation helps in propagation of the signal in an energy efficient way to the functional domain where movement is mainly restricted to the side chains, loops and linker regions and which occur within picosecond to nanosecond timescales [1]. PDZ (postsynaptic density-95/discs large/zonula occludens-1) domains that are involved in myriads of protein-protein interactions [2,3] exhibit minimal structural changes during allosteric propagation. These domains have multiple ligand docking sites and are known to possess unique dynamics that regulate conformation of the functional site from a distal region. HtrA2 (High temperature requirement protease A2), a PDZ bearing protein, is a mitochondrial trimeric pyramidal proapoptotic serine protease with complex domain architecture whose activity is likely regulated by interdomain crosstalk and structural plasticity [4]. Mature HtrA2 comprises 325 amino acids with residues S173, D95 and H65 forming the catalytic triad which is?buried 25 A above the base of the pyramid suggesting requirement of conformational changes for its activation. Apart from PDZ, this multidomain protein has a short N-terminal region, a serine protease domain and a non-conserved flexible linker at the PDZprotease interface [4]. HtrA2 is involved in both caspase dependent as well as caspase independent apoptotic pathways [5,6,7]. Literature suggests it might have chaperoning functions as well and recently has been found to be associated with several neurodegenerative disorders [8,9,10]. Based on information from literature [4,11], this multitasking ability of HtrA2 can be attributed to its serine protease activity which is intricately coordinated by its unique substrate binding Sermorelin site process, complex trimeric structure, interdomain networking and conformational plasticity. However, the unbound inactive form.S, the measurement of overall level of carbonyls of plasma proteins may be used as a surrogate marker of oxidative modification of VWF, as shown in the present study. In conclusion, the emerging scenario shows that the oxidative stress in diabetes involves also VWF and is associated with increased presence of UL-VWF multimers that are involved in the genesis of major cardiovascular events in this clinical setting.AcknowledgmentsRDC thanks prof. V. De Filippis (University of Padua, Italy) for valuable suggestions and scientific discussions. The generous gift of recombinant VWF by Dr. Friedrich Scheiflinger (Baxter Innovations GmbH, Vienna, Austria) is gratefully acknowledged.Author ContributionsEnrolled patients and analyzed the data: FZ. Enrolled patients: FM. Analyzed the data and revised the manuscript: GG. Conceived and designed the experiments: RDC SL DP. Performed the experiments: LO SL RDC PR. Analyzed the data: RDC SL LO DP. Contributed reagents/ materials/analysis tools: RDC SL LO. Wrote the paper: RDC.
Multidomain proteins due to their structural complexity require different levels of regulatory mechanisms for executing cellular functions efficiently within a specified time period. Allosteric modulation of conformations is one such mechanism which often helps a protein to regulate a functional behaviour such as for an enzyme to attain an active functional state upon ligand or substrate binding. In allostery, sometimes there are large conformational changes that require significant rotations and translations of individual domains at the timescales of microsecond to millisecond. While in some other cases, minimal structural perturbation helps in propagation of the signal in an energy efficient way to the functional domain where movement is mainly restricted to the side chains, loops and linker regions and which occur within picosecond to nanosecond timescales [1]. PDZ (postsynaptic density-95/discs large/zonula occludens-1) domains that are involved in myriads of protein-protein interactions [2,3] exhibit minimal structural changes during allosteric propagation. These domains have multiple ligand docking sites and are known to possess unique dynamics that regulate conformation of the functional site from a distal region. HtrA2 (High temperature requirement protease A2), a PDZ bearing protein, is a mitochondrial trimeric pyramidal proapoptotic serine protease with complex domain architecture whose activity is likely regulated by interdomain crosstalk and structural plasticity [4]. Mature HtrA2 comprises 325 amino acids with residues S173, D95 and H65 forming the catalytic triad which is?buried 25 A above the base of the pyramid suggesting requirement of conformational changes for its activation. Apart from PDZ, this multidomain protein has a short N-terminal region, a serine protease domain and a non-conserved flexible linker at the PDZprotease interface [4]. HtrA2 is involved in both caspase dependent as well as caspase independent apoptotic pathways [5,6,7]. Literature suggests it might have chaperoning functions as well and recently has been found to be associated with several neurodegenerative disorders [8,9,10]. Based on information from literature [4,11], this multitasking ability of HtrA2 can be attributed to its serine protease activity which is intricately coordinated by its unique substrate binding process, complex trimeric structure, interdomain networking and conformational plasticity. However, the unbound inactive form.
