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Characterization of oxyanion hole mutants of the cysteine proteases papain and cathepsin B ** check pdfCarrier, Julie, 1959- January 1992 (has links)
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Redox properties of cathepsin B in relation to its activity in vivo.Pillay, Ché Sobashkar. 21 October 2013 (has links)
The main site for protein degradation along the endosomal pathway is believed to be the late
endosome. Lysosomes are thought to be storage organelles that, when necessary, inject
proteases into the late endosome. It was hypothesised that differences in the lumenal redox
environments between the two organelles could be responsible for their functional
differences. In an attempt to quantify this potential difference, the lysosomal cysteine
protease cathepsin B was isolated by an improved purification procedure. Several
intracellular reducing agents were used to activate cathepsin B, the most effective being
cysteine. Cysteine was used to activate cathepsin B under various pH conditions in order to
model endosomal conditions. An inverse relationship was found between the pH and the
concentration of cysteine required to activate cathepsin B. This suggested that cathepsin B
may have an optimal redox potential. In order to determine this potential, cysteinexystine
redox buffers were made up and used in determination of the activity of the enzyme against a
synthetic and a whole protein substrate (haemoglobin). No distinct redox potential could be
determined using either substrate, but it was found that cystine stimulated proteolysis of
haemoglobin. A similar stimulatory effect was observed for cathepsin D and papain
hydrolysis of haemoglobin. This effect is possibly due to the ability of cystine to promote
substrate structure, effectively increasing the substrate concentration. These findings and
other results obtained from the literature have been used to create a model of how proteolysis
may be regulated along the endosomal system. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1999.
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Možnosti a limity RNA interference u klíštěte \kur{Ixodes ricinus} / Potentials and limits of RNA interference in the tick \kur{Ixodes ricinus}MUSIL, František January 2009 (has links)
The function of chitin binding protein (CBP) and two isoforms of cathepsin B (cathB1, cathB2) were tested by using RNA interference in the tick I. ricinus. Two different methods have been used to deliver dsRNA for RNAi in ticks {--} injection and capillary feeding. The synthesized dsRNA was used to find out the impact of RNAi in the tick tissues, which were tested by RT-PCR and Western blot. The expression of CBP was successfully silenced by RNAi in the salivary glands. The silencing of cathB1 and cathB2 in the gut was less effective, but still limited tick`s ability to feed.
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A study of the role of redox potential in lysosomal function.Meinesz, Richard Edward. 11 October 2013 (has links)
No abstract available. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1996.
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Induction of auto-antibodies to Cathepsin B.Moolman, Lizette. 08 November 2013 (has links)
Because tumours are comprised of "self" cells and antigens, they escape recognition by the immune system, which discriminates between "self" and "non-self". One such antigen is cathepsin B, a lysosomal cysteine proteinase, that has been implicated as one of the proteolytic enzymes involved in tumour invasion and metastasis. Cathepsin B autoantibodies
could open possibilities which may be useful in cancer immunotherapy. In this study generation of cathepsin B autoantibodies was attempted by manipulating the immune system into recognising and responding to cathepsin B in complex with a "foreign" protein, bovine serum albumin (BSA). Cathepsin B was isolated from rabbit liver using the three phase partitioning (TPP) method, modified by adding t-butanol in the homogenisation buffer. Isolation of cathepsin Band cathepsin L, using this novel method, minimised the formation of artefacts such as a covalent
cathepsin L-stefin B complex and produced higher yields of enzyme.
Pure rabbit liver cathepsin B was conjugated to BSA, using glutaraldehyde as coupling agent, and administered intramuscularly into rabbits. Another three inoculation protocols, which functioned as controls were: i) free cathepsin B administered intramuscularly, ii) complexed cathepsin B administered intravenously, and iii) free cathepsin B administered intravenously. IgGs isolated from inoculated rabbits' serum were assayed by a three layer ELISA system, immunoinhibition assays and dot blots. The anti-complex (intramuscular) antibodies showed the highest recognition for cathepsin B and were the only antibodies that were immunoinhibitory. This suggests that the immune system was, to some extend, successfully
manipulated into recognising the complexed "self" cathepsin B. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2001.
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Untersuchung zur Funktion von Cathepsin B in der durch zytotoxische T-Zellen vermittelten Lyse von Tumorzellen / Investigation of the function of cathepsin B in T cell mediated tumor cell lysisEnsslen, Miriam 03 August 2009 (has links)
No description available.
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PREPARATION AND EVALUATION OF PEPTIDYL ACYLOXYMETHYL KETONES FOR CATHEPSIN B IMAGINGEdem, Patricia 10 1900 (has links)
<p>This thesis describes the initial steps towards the use of dipeptidyl acyloxymethyl ketones as a platform to develop molecular imaging (MI) probes for cancer. Initially the synthesis of an AOMK was performed following a literature procedure which resulted in an epimerized product. This issue was addressed by optimizing an alternative method yielding all intermediates in yields similar or better to those reported in the literature (final product yield of 67%). An AOMK derivative that can be used to evaluate target expression levels was synthesized by linking a fluorescent dye to the ε-amine group of lysine in accordance to a literature procedure describing the synthesis of an optical imaging probe in 24% yield. A second generation derivative AOMK was prepared by linking 4-fluoro-benzoic acid to the same amino group yielding a model of a PET MI probe.</p> <p>An endpoint colorimetric assay was developed and optimized to test cathepsin B inhibitors. Due to the fact that the AOMKs exhibit time-dependent inhibition these assay conditions did not prove to be adequate for the assessment of the cathepsin B binding. Steps toward developing a continuous assay that would be better suited for these compounds were achieved. Factors such as the relationship between the formation of the assay product vs enzyme concentration and determination of the Michelis-Menten constant (K<sub>m</sub> = 390 ± 30 nM) were established. These parameters can be used to determine the optimal enzyme and substrate concentration that should be used to test the AOMK based probes.</p> / Master of Science (MSc)
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Rôle du CD40 dans la mort cellulaireJundi, Malek January 2008 (has links)
Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.
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Rôle du CD40 dans la mort cellulaireJundi, Malek January 2008 (has links)
Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal
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Characterization of cathepsin b mrna and protein expression, enzymatic activity and cellular localization following contusion spinal cord injury in ratsEllis, Rebecca Catherine, January 2004 (has links)
Thesis (Ph.D.)--University of Florida, 2004. / Typescript. Title from title page of source document. Document formatted into pages; contains 97 pages. Includes Vita. Includes bibliographical references.
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