No description available.

Thesis (Ph. D.)--University of Virginia, 1997. / Spine title: Role of Tryptophan in cytochrome b₅. Includes bibliographical references (187-196). Also available online through Digital Dissertations.

Thesis (Ph.D.) --University of Montana, 2009. / Title from author supplied metadata. Description based on contents viewed on June 11, 2009. ETD number: etd-03252009-151239. Author supplied keywords: Protein Folding ; Denatured State ; Random Coil ; Aggregation ; TR-FRET. Includes bibliographical references.

Thesis (M.S.)--Virginia Polytechnic Institute and State University, 1993. / Vita. Abstract. Includes bibliographical references (leaves 53-69). Also available via the Internet.

No description available.

Streptomyces roseochromogenes, NCIB 10984, hydroxylates exogenous progesterone to 16a hydroxyprogesterone and thereafter in a second phase bioconversion to 2ß, 16a-dihydroxyprogesterone. Characterisation of this reaction was carried out at the whole cell level. The cellular components responsible for this reaction were also purified to homogeneity. S. roseochromogenes contains a cytochrome P450 and two electron transfer proteins, roseoredoxin and roseoredoxin reductase. A reconstituted incubation containing these purified proteins and the natural electron donor, NADH. produced identical hydroxyprogesterone metabolites as intact cells. In sodium periodate (Na104) supported incubations, the initial rate of progesterone hydroxylation was marginally higher than in the natural reconstituted system but the product yield was significantly lower. The yield data showed that the reconstituted natural pathway, supported multiple rounds of hydroxylation in contrast to a likely single round by a minority of P450s in the periodate reaction. When S. roseochromogenes was incubated with exogenous progesterone for 25 h the major metabolite, 16a-hydroxyprogesterone was produced in 3.6 fold excess to the minor metabolite 2ß, 16a-dihydroxyprogesterone. In a reconstituted system containing highly purified progesterone 16a-hydroxylase cytochrome P450, roseoredoxin and roseoredoxin reductase, both metabolites were produced but in a 10: 1 ratio. When S. roseochromogenes was preincubated with progesterone and the purified components of the hydroxylase system assayed as before, the ratio of 16a-hydroxyprogesterone to 2ß, 16adihydroxyprogesterone produced, decreased to 2.8: 1, virtually identical to the ratio in whole cell biotransformations. Reconstitution assays containing all combinations of hydroxylase proteins purified from progesterone preincubated and control cells, identified roseoredoxin as solely responsible for the observed changes in in vitro metabolite ratios. The fact that the 2.8: 1 ratio was also obtained when S. roseochromogenes was exposed to cycloheximide prior to progesterone pre-incubation; pointed to post translation modification of roseoredoxin. Separation of two isoforms by 2-D isoelectric focusing supported this proposition. A partial 10 amino acid sequence was obtained for both the cytochrome P450 and roseoredoxin for the purpose of probe design for eventual cloning. An amino acid sequence search revealed this P450 to be unique and unlike any other known P450 sequence. These two proteins were also successfully crystallised by hanging drop vapour diffusion trials, giving isomorphous crystals. These crystals will be used for structure determinations pending further growth.

Thèse de doctorat : Biologie moléculaire végétale : Strasbourg 1 : 2008. / Titre provenant de l'écran-titre. Bibliogr. p. 126-140.

The mitochondria are the primary energy providers for most eukaryotic cells. The substrate and products of the mitochondria need to be translocated across the semi-permeable mitochondrial outer membrane (MOM). Mitochondrial porin is an aqueous channel in the MOM thought to provide the primary pathway for metabolite translocation. Porin is a nuclear encoded protein and therefore needs to be transported to the mitochondria, translocated across and assembled within the MOM. Of all the recognition signals required for successful transport, import and assembly, only the β-sorting signal used in assembly is known. In addition, this protein possesses the ability to gate, and in doing so can preferentially allow the passage of anions in the open state and cations in the closed state. However, the precise mechanism by which gating of porin occurs and a complete understanding of porin’s function in vivo remains elusive. The essentiality of porin was examined by constructing a strain of Neurospora crassa deficient for porin. This strain, denoted as WS004, exists as evidence that porin is non-essential for the survival of Neurospora crassa. However, the loss of porin results in a reduction in growth rate due to the dysfunction of the cytochrome mediated respiratory pathway, which was made evident by the reduction of cytochrome b and almost complete lack of cytochrome aa3. WS004 survives by inducing the expression of alternative oxidase, which funnels the electrons from the Q pool directly to oxygen, bypassing the cytochrome b and aa3 containing complexes III and IV respectively. Additional phenotypic differences observed included loss in ability to produce aerial hyphae, reduced amount of conidia produced and strains that were female sterile. It was determined, that additional genetic factors influenced the resulting phenotype due to the loss of porin. LC-MS/MS, in combination with iTRAQ labelling, was utilized to examine changes in the proteome profiles of porin containing and porin lacking mitochondria and showed several different proteins as significantly up- or down-regulated which lend to an explanation to some of the phenotypes observed. Taken together, these results demonstrate the central role of porin in regulating both mitochondrial and cellular processes.

The mitochondria are the primary energy providers for most eukaryotic cells. The substrate and products of the mitochondria need to be translocated across the semi-permeable mitochondrial outer membrane (MOM). Mitochondrial porin is an aqueous channel in the MOM thought to provide the primary pathway for metabolite translocation. Porin is a nuclear encoded protein and therefore needs to be transported to the mitochondria, translocated across and assembled within the MOM. Of all the recognition signals required for successful transport, import and assembly, only the β-sorting signal used in assembly is known. In addition, this protein possesses the ability to gate, and in doing so can preferentially allow the passage of anions in the open state and cations in the closed state. However, the precise mechanism by which gating of porin occurs and a complete understanding of porin’s function in vivo remains elusive. The essentiality of porin was examined by constructing a strain of Neurospora crassa deficient for porin. This strain, denoted as WS004, exists as evidence that porin is non-essential for the survival of Neurospora crassa. However, the loss of porin results in a reduction in growth rate due to the dysfunction of the cytochrome mediated respiratory pathway, which was made evident by the reduction of cytochrome b and almost complete lack of cytochrome aa3. WS004 survives by inducing the expression of alternative oxidase, which funnels the electrons from the Q pool directly to oxygen, bypassing the cytochrome b and aa3 containing complexes III and IV respectively. Additional phenotypic differences observed included loss in ability to produce aerial hyphae, reduced amount of conidia produced and strains that were female sterile. It was determined, that additional genetic factors influenced the resulting phenotype due to the loss of porin. LC-MS/MS, in combination with iTRAQ labelling, was utilized to examine changes in the proteome profiles of porin containing and porin lacking mitochondria and showed several different proteins as significantly up- or down-regulated which lend to an explanation to some of the phenotypes observed. Taken together, these results demonstrate the central role of porin in regulating both mitochondrial and cellular processes.

Thesis (Ph. D.)--Ohio State University, 2008. / Title from first page of PDF file. Includes bibliographical references (p. 113-119).