Mitochondria, analysis

It is important to appreciate that this principle of coupling-in-series underlies all biochemical pathways or processes, e.g. glycolysis, generation of ATP in the mitochondrion, protein synthesis from amino acids or a signal transduction pathway. Indeed, despite the fundamental importance of signalling pathways in biochemistry, a thermodynamic analysis of such a pathway has never been done, but the principles outlined above must apply even to signalling pathways. 

Petry F, Harris JR (1999) Ultrastructure fractionation and biochemical analysis of Cryptosporidium parvum sporozoites. Int J Parasitol 29 1249-1260 Putignani L, Tait A, Smith HV, Horner D, Tovar J, Tetley L, Washing JM (2004) Characterization of a mitochondrion-like organelle in Cryptosporidium parvum. Parasitol 129 1-18... 

Haresh K, Suresh K, Khairul Anus A, Saminathan S (1999) Isolate resistance of Blastocystis hominis to metronidazole. Trop Med Int Health 4 274-277 Inui H, Ono K, Miyatake K, Nakano Y, Kitaoka S (1987) Purification and characterization of pyruvate NADP+ oxidoreductase in Euglena gracilis. J Biol Chem 262 9130-9135 Keithly JS, Langreth SG, Buttle KF, Mannella CA (2005) Electron tomographic and ultra-structural analysis of the Cryptosporidium parvum relict mitochondrion, its associated membranes, and organelles. J Eukaryot Microbiol 52 132-140 Kurland CG, Andersson SGE (2000) Origin and evolution of the mitochondrial proteome. Micro Mol Biol Rev 64 786-820... 

Keithly JS, Langreth SG, Buttle KF, Mannella CA (2005) Electron tomographic and ultrastruc-tural analysis of the Cryptosporidium parvum relict mitochondrion, its associated membranes and organelles. J Eukaryot Microbiol 52 132-140 Knight J (2004) Giardia not so special, after all Nature 429 236-237... 

This chapter starts with a simplified analysis of biological processes using the basic tools of physics, chemistry, and thermodynamics. It provides a brief description of mitochondria and energy transduction in the mitochondrion. The study of proper pathways and multi-inflection points in bioenergetics are summarized. We also summarize the concept of thermodynamic buffering caused by soluble enzymes and some important processes of bioenergetics using the linear nonequilibrium thermodynamics formulation. 

Proteins are part of a dynamic network of biomolecules that interact to regulate their localization and function within the cell. Disruption of this physical and chemical system of interactions has become the first paramount step in performing protein analysis by shotgun proteomics. Protein isolation techniques, for instance, have allowed understanding of the complex dynamics of proteins among cellular subcompartments, such as the nucleolus or the mitochondrion (6). Membrane-embedded proteins (7) and DNA-binding transcription factors (8) are two other prominent examples where inadequate protein extraction may hamper further analysis by LC-MS. 

Q.2.2 Extend your systems analysis to various subsystems of the cell including (1) the cytosol, (2) the ribosome (3) mitochondrion (4) cell membrane (5) nucleus. 

In these situations, one approach has been to perform further testing using mtDNA sequencing. Since there are multiple copies of mitochondrion in each cell, the likelihood of obtaining a result is greatly increased when compared to nuclear DNA. However, mtDNA analysis involves difficult and expensive analytical procedures, and because it is inherited solely through the female fine, maternally related individuals will all have the same profile and statistical results are much less conclusive. 

The method of estimating the amount of inner membrane in mitochondria has varied from investigation to investigation. In a few cases morphometric analysis of the amount of membrane per square micrometer of mitochondrial profile has been made in some, inner membrane is measured in terms of the phospholipid or cardiolipin content of the mitochondrion, the latter being particularly suitable since cardiolipin is fairly well localized in the inner membrane of mitochondria in others, mitochondria have been disrupted by osmotic shock, detergents, and sonication, and the protein content of the insoluble fraction has then been... 

From a biophysical point of view, the use of whole mitochondrion (or chloroplast) to test the "molecular" theory of Mitchell does not seem satisfying. The translocation of ions, such as protons, across a complex system, such as a cristae, is in itself ill-defined. The interactions between various fluxes (ion and water movements, electron and hole transport, etc.) are far too complex to be amenable to a simple analysis. At the present time, direct tests with the mitochondrial membranes are difficult. Experimental testing of the chemiosmotic hypothesis, using simpler model systems such as planar BLM and spherical liposomes are, therefore, in order. 

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