The 18 kDa translocator protein (TSPO) is an integral membrane protein known to be involved in porphyrin transport and heme biosynthesis. TSPO also plays a role in modulating immune response, regulation of cell proliferation and programmed cell death. TSPO is associated with several health conditions such as ischemic heart reperfusion, cancers and neurodegenerative diseases. While the potential of TSPO as a major therapeutic target has been well recognized, the lack of detailed understanding of its function and ligand binding sites have hindered drug design. In this thesis we will describe the expression, extraction and purification of Rhodobacter sphaeroides TSPO (RsTSPO). Rhodobacter is one of the closest ancestors of mitochondria and RsTSPO can be used as a model for mammalian TSPO. RsTSPO was overexpressed in E. coli, and it was extracted from the membranes with detergent (i.e. n-dodecylphosphocoline (DPC) and n-dodecyl-ß-D-maltoside (DDM)) then purified by Ni-affinity and size exclusion chromatographies. The RsTSPO could be expressed and purified in high yield (mg quantitites), high purity, and in its functional state. Binding studies of RsTSPO were performed utilizing fluorescence quenching of intrinsic tryptophans with a number of porphyrins ligands such as protoporphyrin IX (PPIX), hemin, meso-Tetra(4-sulfonatophenyl)porphine dihydrochloride (2HTPPS), Zn (II) meso-Terta(4-sulfonatophenyl)porphine dihydrochloride (ZnTPPS) and with the diagnostic ligand, PK11195. Binding studies of RsTSPO were performed in both detergents (DPC and DDM) to assess the effect of the membrane mimic environment on binding. We will also describe the first evidence of the catalytic activity of the purified RsTSPO under ambient light in the presence of molecular oxygen on its natural ligand, protoporphyrin IX. We will demonstrate the photodegradation of protoporphyrin IX by RsTSPO in the presence of oxygen and light. In addition, we have studied the effect of molecular oxygen and reactive oxygen scavengers (sodium azide and glutathione) on the decomposition of protoporphyrin IX. Also, we will describe preliminary NMR studies in an effort to characterize the ligand binding site(s) of TSPO. We have performed both solution and solid state NMR studies on pure RsTSPO and RsTSPO-ligand complexes.

This book is a printed edition of the Special Issue "Translocator Protein (TSPO)" that was published in IJMS

Plant cells house highly dynamic cytoskeletal networks of microtubules and actin microfilaments. They constantly undergo remodeling to fulfill their roles in supporting cell division, enlargement, and differentiation. Following early studies on structural aspects of the networks, recent breakthroughs have connected them with more and more intracellular events essential for plant growth and development. Advanced technologies in cell biology (live-cell imaging in particular), molecular genetics, genomics, and proteomics have revolutionized this field of study. Stories summarized in this book may inspire enthusiastic scientists to pursue new directions toward understanding functions of the plant cytoskeleton. The Plant Cytoskeleton is divided into three sections: 1) Molecular Basis of the Plant Cytoskeleton; 2) Cytoskeletal Reorganization in Plant Cell Division; and 3) The Cytoskeleton in Plant Growth and Development. This book is aimed at serving as a resource for anyone who wishes to learn about the plant cytoskeleton beyond ordinary textbooks.

This volume of International Review of Neurobiology concentrates on modern concepts of focal epileptic networks. The volume addresses specific topics such as seizures (including transition and termination), limbic networks, alteration of metabolism, and neocortical focus and malformation of cortical development, among others. Published since 1959, International Review of Neurobiology is a well-known series appealing to neuroscientists, clinicians, psychologists, physiologists, and pharmacologists. Led by an internationally renowned editorial board, this important serial publishes both eclectic volumes made up of timely reviews, and thematic volumes that focus on recent progress in a specific area of neurobiology research. Our knowledge about the mechanisms involved in pathophysiology of epilepsy has rapidly expanded during last decade This special volume brings overview about modern concepts of epileptic focus organization and about the altered neural network dynamics which results in propensity of the brain tissue to generate spontaneous and repeated seizures

Coronary Microvascular Obstruction in Acute Myocardial Infarction: From Mechanisms to Treatment provides a comprehensive understanding of the phenomenon of coronary microvascular obstruction (CMVO) that is the main limitation of reperfusion therapies in ST-elevation myocardial infarction. It provides in-depth coverage of the phenomenon of CMVO which heavily affects prognosis by increasing the risk of death and heart failure at follow-up. A first of its kind reference dedicated solely to this topic, it is appropriate for a wide audience, from researchers, to those who aid in the management, prevention and treatment of CMVO. Provides in-depth coverage of coronary microvascular obstruction (CMVO), spanning research, management, prevention and treatment Includes the most up-to-date information on CMVO as presented from top experts around the word Provides access to a companion website with extra material, including tables, additional references and instructional videos Gives extensive coverage on how to measure CMVO, including in-depth indexes that can be used to detect and quantify the phenomenon

Psychotic Disorders: Comprehensive Conceptualization and Treatments emphasizes a dimensional approach to psychosis--one of the most fascinating manifestations of altered brain behavior--that cuts across a broad array of psychiatric diagnoses from schizophrenia to affective psychosis and organic disorders like epilepsy and dementias. Written by an international roster of over seventy leading experts in the field, this volume comprehensively reviews, critiques, and integrates available knowledge on the etiology, mechanisms, and treatments of psychotic disorders, and outlines ways forward in both research and clinical practice towards more objective, mechanistically-based definitions of psychotic disorders. Chapters address topics such as psychosis phenomenology, biomarkers and treatments, the overlaps and interfaces between psychiatric disorders within the psychosis dimension, and novel disease definitions. Furthermore, the volume incorporates findings on potential mechanisms, bridges between various system levels (i.e., genetic, epigenetic, molecular and cellular, brain circuit and function, psychological, social, environmental and cultural) and their interactions, as well as the potential role in causation and/or mediation in psychotic disorders. Finally, the volume outlines a broad array of treatment approaches, from the readily available (e.g., psychopharmacology, various modalities of psychotherapy) to the experimental (e.g., cognitive interventions, neuromodulation). With a concluding section of forward perspectives conjecturing future directions and related challenges, this book aspires to stimulate new knowledge, generate novel frameworks, and carry new directions forward on psychotic disorders.