No description available.

Thesis (Ph. D.)--Virginia Commonwealth University, 2006. / Prepared for: Dept. of Anatomy and Neurobiology . Bibliography: leaves 108 - 125. Also available online.

This study had two aspects. The first was the production and purification of TIMP-l. The second was a series of biophysical studies of TIMP-l and a TIMP-l derived peptide. A monoclonal antibody affinity column was developed and used to purifY large quantities of human TIMP-l for further experiments. Two E.coli expression systems were studied to determine whether they would be suitable for large scale production of recombinant protein. In the first system TIMP-I was to be secreted as a fusion protein which could be cleaved, leaving a free N-terminus. It was discovered that it was not possible to cleave off the fusion protein. In the second system, the protein was secreted, without additions to the periplasm. Although active protein, with the correct N-tenninus, was obtained, the yields were too low to be of use for large scale expression. Secondary structure analysis by CD and FTIR showed TIMP-l to be a mostly f3- sheet protein (approaching 50%) with around 20% a-helix. A temperature study using these techniques found that little change occurs until temperatures of over 60°C where the protein aggregates. The small changes appear to be a general loosening of the structure. In analyses of the surface of TIMP-l, additional carbohydrate was identified (other than the two N-linked chains) using Con-A probing of Western blots. TIMP-l purified from WI-38 foetal lung fibroblast cells can be separated into two pools by Concanavalin A-Sepharose chromatography. These two pools were found to have a different set of pIs and a different monosaccharide composition. The use of NMR paramagnetic probes identified a hydrophobic region exposed on the surface of TIMP-I. This region probably includes a tyrosine residue and either a tryptophan or phenylalanine. The presence of an exposed hydrophobic region was also shown in binding studies using the fluorescent probe ANS. These studies identified a single, low affinity binding site. An additional study with the N-terminal fragment of type-I collagenase found no binding sites on the enzyme, but a change in fluorescence occurred when TIMP-I was present. A peptide was designed based on the N-terminal sequence of TIMP-I. High homology, susceptibility to mutation and an interesting resemblance to the Bowman-Birk family of inhibitors suggested that this peptide might be inhibitory. It was found to have only a weak inhibitory activity against gelatinase. NMR studies of this peptide in water showed a large number of conformers as a result of stabilisation of the cis isomer of its proline residues. This preference for the cis form was retained for one proline in the solvent, TFE. Preliminary NMR studies were also carried out which concluded that TIMP-I should be suitable for further structural studies using isotopic labelling.

Orientador: Ana Paula de Souza Pardo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-15T12:46:18Z (GMT). No. of bitstreams: 1 Planello_AlineCristiane_M.pdf: 940083 bytes, checksum: 36ed3b0b3b0d10c144805116e08917dd (MD5) Previous issue date: 2010 / Resumo: Objetivo: As Metaloproteinases da Matriz ( MMPs) são enzimas que degradam a matriz extracelular (MEC) e tem sido associadas às desordens temporomandibulares (DTM). Nós investigamos a freqüência dos -1607 1G/2G MMP1 polimorfismo (rs1799750), -1171 6A/5A MMP3 polimorfismo (rs3025058) e -1562 C/T MMP9 polimorfismo (rs3918242) em indivíduos com sinais de degeneração da ATM, diagnosticados por exame de imagem, a fim de analisar a associação desses polimorfismos e a DTM. Métodos: A população estudada foi composta por 115 indivíduos diagnosticados por exame de imagem (grupo DTM) e 117 controles. Os polimorfismos genéticos foram determinados por PCR/RFLP. Resultados: A freqüência do genótipo 2G/2G no gene MMP1 foi significantemente mais alta no grupo DTM do que no grupo Controle (p = 0.008). O genótipo 2G/2G no grupo DTM mostrou um risco aumentado para a DTM com um OR = 2.25 ( 95% IC = 1.26 - 3.99) quando comparado com os genótipo 1G/2G e 1G/1G. A freqüência dos alelos do gene MMP1 não mostrou diferença significativa entre os grupos (p > 0.05). A distribuição dos genótipos e alelos dos genes MMP3 e MMP9 não mostrou diferença significativa (p > 0.05). Conclusão: Nossos resultados mostram a associação entre o polimorfismo -1607 MMP1 e a suscetibilidade à DTM / Abstract: Objective. Matrix metalloproteinases (MMPs) degrade extracellular matrix components and have been implicated to play an important role in temporomandibular joint disorder (TMD). We investigated the frequency of -1607 1G/2G MMP1 polymorphism (rs1799750), -1171 6A/5A MMP3 polymorphism (rs3025058) and -1562 C/T MMP9 polymorphism (rs3918242) in individuals with TMJ degeneration diagnosed by image exam in order to analyze the association of these MMPs polymorphisms and TMD. Methods. The studied population comprised 115 TMD individuals diagnosed by image exam and 117 healthy controls. Genotypes were determined using polymerase chain reaction/Restriction fragment length polymorphism PCR/RFLP. Results. The MMP1 2G/2G genotype was significantly higher in the TMD group than in the Control group (p = 0.008). The genotype 2G/2G in the TMD group showed an increased risk to TMD with an OR = 2.25 (95% CI = 1.26 - 3.99) when compared with 1G/2G and 1G/1G genotypes. Analysis of MMP1 allele frequencies showed no significant difference (p > 0.05). The MMP3 and MMP9 genotypes distribution and alleles frequency did not differ between the groups (p > 0.05). Conclusion. Our results report the association of -1607 MMP1 gene polymorphism and increased risk to TMD / Mestrado / Histologia e Embriologia / Mestre em Biologia Buco-Dental

No description available.

Reproduction de : Thèse doctorat : Médecine. Biomolécules et dynamique cellulaire : Reims : 2006. / Titre provenant de l'écran-titre. Bibliogr. p.110-148.

Reproduction de : Thèse doctorat : Pharmacie. Sciences du médicament : Reims : 2008. / Titre provenant de l'écran-titre. Bibliogr.

Thesis (M.S.)--University of Michigan, 2003. / Includes bibliographical references.

Matrix Metalloproteinase-13 (MMP-13) belongs to a large family of proteolytic enzymes which are characterized by their ability to degrade the extracellular matrix components. MMP-13 appears to have a critical role in tumor invasion and metastasis. In this study, several fluorogenic probes specific for MMP-13 were designed and characterized. These synthesized probes could be modified with chelators to be applied for imaging MMP-13 in breast cancer and/or multiple myeloma models. The activity and selectivity of MMP-13 and other MMPs against these probes were studied through two approaches. It was found that these probes were cleaved by all MMPs, but MMP-13 showed the highest activity and selectivity towards these peptides. / Includes bibliography. / Thesis (M.S.)--Florida Atlantic University, 2020. / FAU Electronic Theses and Dissertations Collection

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.