ABSTRACT
Peptides hold great potential for the cancer therapy and diagnostics. In the current review, the main and most influential areas of peptide cancer therapeutics are overviewed. These include the development of anticancer peptides, use of peptides for drug delivery, and cancer targeting.
Subject(s)
Antineoplastic Agents/administration & dosage , Drug Delivery Systems , Neoplasms/drug therapy , Peptides/administration & dosage , Animals , Humans , Molecular Targeted TherapyABSTRACT
Aging-related oxidative stress has been linked to degenerative modifications in different organs and tissues. Using redox proteomic analysis and illustrative tandem mass spectrometry mapping, we demonstrate oxidative posttranslational modifications in structural proteins of intervertebral discs (IVDs) isolated from aging mice. Increased protein carbonylation was associated with protein fragmentation and aggregation. Complementing these findings, a significant loss of elasticity and increased stiffness was measured in fibrocartilage from aging mice. Studies using circular dichroism and intrinsic tryptophan fluorescence revealed a significant loss of secondary and tertiary structures of purified collagens following oxidation. Collagen unfolding and oxidation promoted both nonenzymatic and enzymatic degradation. Importantly, induction of oxidative modification in healthy fibrocartilage recapitulated the biochemical and biophysical modifications observed in the aging IVD. Together, these results suggest that protein carbonylation, glycation, and lipoxidation could be early events in promoting IVD degenerative changes.
Subject(s)
Aging/metabolism , Fibrocartilage/metabolism , Intervertebral Disc Degeneration/metabolism , Intervertebral Disc Degeneration/pathology , Intervertebral Disc/metabolism , Intervertebral Disc/pathology , Protein Carbonylation , Amino Acid Sequence , Animals , Biomechanical Phenomena , Collagen/chemistry , Collagen/metabolism , Intervertebral Disc/physiopathology , Intervertebral Disc Degeneration/physiopathology , Male , Matrix Metalloproteinases/metabolism , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Oxidative Stress , Protein Folding , Protein Multimerization , Protein Structure, Quaternary , ProteolysisABSTRACT
Lipid modifications are essential in cellular sorting and trafficking inside cells. The role of phosphoinositides in trafficking between Golgi and endocytic/lysosomal compartments has been extensively explored and the kinases responsible for these lipid changes have been identified. In contrast, the mechanisms that mediate exit and recycling from lysosomes (Lys), considered for a long time as terminal compartments, are less understood. In this work, we identify a dynamic association of the lipid kinase PI4KIIIß with Lys and unveil its regulatory function in lysosomal export and retrieval. We have found that absence of PI4KIIIß leads to abnormal formation of tubular structures from the lysosomal surface and loss of lysosomal constituents through these tubules. We demonstrate that the kinase activity of PI4KIIIß is necessary to prevent this unwanted lysosomal efflux under normal conditions, and to facilitate proper sorting when recycling of lysosomal material is needed, such as in the physiological context of lysosomal reformation after prolonged starvation.
Subject(s)
1-Phosphatidylinositol 4-Kinase/metabolism , Lipid Metabolism , Lysosomal-Associated Membrane Protein 1/metabolism , Lysosomes/metabolism , Lysosomes/physiology , Animals , Biological Transport/physiology , COS Cells , Chlorocebus aethiops , Gene Knockdown Techniques , HEK293 Cells , Humans , Immunohistochemistry , Lentivirus , Lysosomes/ultrastructure , Male , Mice , Mice, Inbred C57BL , Microscopy, Electron , Microscopy, Fluorescence , NIH 3T3 Cells , Protein Isoforms/metabolism , RNA, Small Interfering/genetics , Rats , Rats, Wistar , Real-Time Polymerase Chain ReactionABSTRACT
In this study a proteomic approach was used to define the protein content of matched samples of afferent prenodal lymph and plasma derived from healthy volunteers. The analysis was performed using two analytical methodologies coupled with nanoliquid chromatography-tandem mass spectrometry: one-dimensional gel electrophoresis (1DEF nanoLC Orbitrap-ESI-MS/MS), and two-dimensional fluorescence difference-in-gel electrophoresis (2D-DIGE nanoLC-ESI-MS/MS). The 253 significantly identified proteins (p<0.05), obtained from the tandem mass spectrometry data, were further analyzed with pathway analysis (IPA) to define the functional signature of prenodal lymph and matched plasma. The 1DEF coupled with nanoLC-MS-MS revealed that the common proteome between the two biological fluids (144 out of 253 proteins) was dominated by complement activation and blood coagulation components, transporters and protease inhibitors. The enriched proteome of human lymph (72 proteins) consisted of products derived from the extracellular matrix, apoptosis and cellular catabolism. In contrast, the enriched proteome of human plasma (37 proteins) consisted of soluble molecules of the coagulation system and cell-cell signaling factors. The functional networks associated with both common and source-distinctive proteomes highlight the principal biological activity of these immunologically relevant body fluids.
