It is generally believed that ribosome-inactivating proteins (RIPs) are transported to their intracellular targets to express their toxicity. However, studies on the uptake, intracellular transport and apoptotic mechanism of type I RIPS and the cell-binding B chain of type II RIPS are rare. This study is to investigate some problems in these aspects of RIP toxicity. / RCA caused a cell loss at the minimal dose of 50 nM at 24 hr. The main type of cell death was apoptosis, which peaked at 12 hr. The apoptosis proceeded through an extrinsic pathway that involved the activation of caspase-8, but not caspase-9. / RTA caused cell loss at the minimal dose of 50 nM at 24 hr. The main type of cell death was apoptosis, which peaked at 12 hr. RTA was internalised via a clathrin-dependent RME. Like the TCS transport, RTA was not found in the Golgi apparatus. The apoptosis proceeded via the extrinsic pathway that involved the activation of caspase-8 and caspase-3. However, on live rabbits, RTA caused necrotic skin damage. / RTB caused cell loss at the minimal dose of 100 nM at 24 hr. The main type of cell death was initially necrosis, but later became apoptosis, which peaked at 12 hr. / TCS caused a decrease in cell number at the minimum effective dose of 800 nM at 24 hr post administration. The main type of cell death was apoptosis, which peaked at 12 hr. / These results show that (1) the cell-binding B chain is not a precondition for RIP toxicity, because TCS and RTA are also toxic to cells; (2) RTB itself is toxic; (3) without the binding of the B chain to cell surface, the entry and intracellular transport of type I RIPS differ from those of the type II; and (4) both RIPs and single B chain can induce apoptosis. Additionally, the results from live rabbits and cultured cells show that in vivo and in vitro toxicity may involve different cell death mechanisms. RTB-treated NIH 3T3 cells may serve as a model for the switch of cell death from necrosis to apoptosis. (Abstract shortened by UMI.) / We studied trichosanthin (TCS) and ricin A chain (RTA), which are type I RIPS, ricinus communis agglutinin (RCA), which is a type II RIP, and ricin B chain (RTB), which is the cell-binding chain of ricin and RCA. / Sha Ou. / "August 2005." / Source: Dissertation Abstracts International, Volume: 67-07, Section: B, page: 3547. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 185-217). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract in English and Chinese. / School code: 1307.

by Mock Wai Yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 112-122). / ACKNOWLEDGEMENTS --- p.I / ABSTRACT --- p.II / TABLE OF CONTENT --- p.IV / Chapter CHAPTER 1: --- INTRODUCTION --- p.1 / Chapter CHAPTER 2: --- PURIFICATION OF α- AND β-MMCs --- p.29 / Chapter CHAPTER 3: --- RIBONUCLEASE ACTIVITY OF MMCs --- p.50 / Chapter CHAPTER 4: --- PURIFICATION AND RIBONUCLEASE ACTIVITY OF RNase-MC --- p.85 / Chapter CHAPTER 5: --- CONCLUSION --- p.109 / REFERENCES --- p.112

Plant cells resistant to specific amino acid analogs have been reported to accumulate the corresponding free amino acids. The purpose of this study was to determine the concentrations of fifteen free amino acids: alanine, valine, leucine, isoleucine, glutamate, proline, arginine, aspartate, threonine, methionine, lysine, serine, glycine, tryptophan and phenylalanine in Daucus carota cell lines, resistant either to the proline analog azetidine-2 carboxylic acid (A2C), or to the tryptophan analog 5-methyltryptophan (5-MT), or to both the analogs combined. This study also intended to determine if these analogs influence the biosynthesis of the above-mentioned fifteen amino acids in the cell line resistant to A2C and 5-MT. Carrot cell lines resistant to 5-MT, to A2C, or to both the analogs were selected by incubating carrot cells in liquid growth media containing either 0.3 mM 5-MT, or 0.5 mM A2C for 6 to 16 weeks. Free amino acid concentrations were then determined in the extracts of the cells. Resistance to 5-MT resulted in significant increases in the intracellular concentrations of tryptophan, phenylalanine, leucine, valine, isoleucine, and proline. Resistance to A2C resulted in significant increase in proline only. Resistance to both the analogs caused increases in proline, lysine, phenylalanine, and tryptophan concentrations. In the cell line resistant to both the analogs, the treatment with 5-MT caused increases in leucine, proline, aspartate, threonine, lysine, and tryptophan. The treatment with A2C caused increases in isoleucine, arginine, threonine, methionine, lysine, and glycine, whereas treatment with both the analogs caused increases in threonine, lysine, phenylalanine, and tryptophan. These results indicate the possibility of a common biosynthetic control of a number of amino acids in carrot cells, resembling that found in microorganisms. It is also evident from the results that the analogs play an active role in the biosynthesis of amino acids in the resistant cell lines.

No description available.

by Chi-Ming Wong. / Bibliography: leaves 183-193 / Thesis (M.Ph.)--Chinese University of Hong Kong, 1984

by Cha Chi Fai. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (p. 139-154). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web.

Further in vivo and in vitro studies using the truncated OsRMR1 proteins from the culture media of transgenic BY-2 cells demonstrated that OsRMR1 functioned as a receptor in transporting vicilin-like storage proteins via specific interaction with their vacuolar sorting determinants. Taken together, the OsRMR1 is a sorting receptor for the PSV pathway that defines a novel organelle as PVC for PSV in rice. / Receptor-mediated protein sorting is one of the mechanisms for transporting soluble proteins to the protein storage vacuoles (PSVs) in plant cells. Members of vacuolar sorting receptor (VSR) family proteins and receptor homology region-transmembrane domain-RING-H2 (RMR) family proteins have been shown to function in mediating the transport of storage proteins to PSVs in plants. However, no prevacuolar compartment (PVC) for the PSV pathway has been identified. In this study, I used a rice RMR protein (OsRMR1) as a probe to study the PSV pathway in rice. Using confocal immunofluorescent and immunogold electron microscopy (EM) with specific OsRMR1 antibodies, I have identified a novel organelle as a PVC for the PSV pathway, because OsRMR1 antibodies labeled the Golgi apparatus, trans-Golgi network (TGN) and the novel organelle in both rice cultured cells and developing rice seeds, as well as the protein body Type II (PBII) in developing rice seeds. This novel organelle is morphologically distinct from the lytic PVC or multivesicular body (MVB). / Shen, Yun. / "May 2008." / Adviser: Liwen Jiang. / Source: Dissertation Abstracts International, Volume: 70-03, Section: B, page: 1428. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (p. 124-139). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Receptor-like protein kinases (RLKs) containing an extracellular leucine-rich-repeat (eLRR) domain, a transmembrane domain, and a cytoplasmic kinase domain play important roles in plant disease resistance. Simple eLRR domain proteins structurally resembling the extracellular portion of the RLKs may also participate in signaling transduction and plant defense response. Yet the molecular mechanisms and subcellular localization in regulating plant disease resistance of these simple eLRR domain proteins are still largely unclear. We provided the first experimental evidence to demonstrate the endosomal localization and trafficking of a novel simple eLRR domain protein (OsLRR1) in the endosomal pathway, using both confocal and electron microscopy. Yeast 2-hybrid and in vitro pull-down assays show that OsLRR1 interacts with the rice hypersensitive induced response protein 1 (OsHIR1) which is localized on plasma membrane. The interaction between LRR1 and HIR1 homologs was shown to be highly conserved among different plant species, suggesting a close functional relationship between the two proteins. The function of OsLRR1 in plant defense response was examined by gain-of-function tests using transgenic Arabidopsis thaliana. The protective effects of OsLRR1 against bacterial pathogen infection were shown by the alleviating of disease symptoms, lowering of pathogen titers, and higher expression of defense marker genes. / Zhou, Liang. / Adviser: Hon Ming Lam. / Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 90-107). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

by Yuan Dingyang. / "September 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 206-232). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Wong Ho Wan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 140-153). / Abstracts in English and Chinese. / Thesis Committee --- p.I / Statement --- p.II / Acknowledgements --- p.III / Abstract --- p.IV / 摘要 --- p.VI / List of Tables --- p.VIII / List of Figures --- p.IX / List of Abbreviations --- p.XI / Table of Contents --- p.XIII / Chapter 1 --- General introduction --- p.1 / Chapter 2 --- Literature reviews --- p.4 / Chapter 2.1 --- LRP and winged bean --- p.4 / Chapter 2.1.1 --- Nutritional values of crop plants --- p.4 / Chapter 2.1.2 --- Lysine-rich protein (LRP) --- p.7 / Chapter 2.1.2.1 --- Identification of lysine-rich protein (LRP) --- p.7 / Chapter 2.1.2.2 --- Cloning cDNA for WBLRP --- p.7 / Chapter 2.1.2.3 --- Transgenic Expression of LRP in other plants --- p.8 / Chapter 2.1.3 --- Unknowns remained --- p.8 / Chapter 2.2 --- Food allergy and gastro-immunity --- p.10 / Chapter 2.2.1 --- What is allergy? 一 A brief introduction --- p.10 / Chapter 2.2.2 --- Food allergy and its symptoms --- p.12 / Chapter 2.2.3 --- Gastrointestinal immunity --- p.13 / Chapter 2.2.4 --- Possible mechanism of food allergy --- p.16 / Chapter 2.2.5 --- Available tests and limitations --- p.18 / Chapter 2.2.6 --- Radioallergosorbent test (RAST) --- p.19 / Chapter 2.2.7 --- Digestibility test --- p.20 / Chapter 2.2.8 --- Betv-1 Allergen Family --- p.21 / Proteins --- p.23 / Chapter 2.3 --- Pathogenesis-related proteins --- p.23 / Chapter 2.3.1 --- Defense-related proteins and pathogenesis-related proteins (PRs) --- p.23 / Chapter 2.3.2 --- Class 10 PR proteins (PR-10s) --- p.25 / Chapter 2.3.3 --- The expression patterns of PR-10s --- p.27 / Chapter 2.3.3.1 --- Pathogens-induced and signal-induced expression --- p.27 / Chapter 2.3.3.2 --- Spatially- and developmentally-regulated expression --- p.28 / Chapter 2.3.3.3 --- Other induction patterns --- p.29 / Chapter 2.3.4 --- Functions ofPR-10s --- p.30 / Chapter 2.4 --- Development of hypotheses and experiments --- p.32 / Chapter 3 --- Materials and methods --- p.36 / Chapter 3.1 --- Introduction --- p.36 / Chapter 3.2 --- Materials --- p.38 / Chapter 3.2.1 --- Chemicals --- p.38 / Chapter 3.2.2 --- Apparatus and commercial kits --- p.39 / Chapter 3.2.3 --- Vectors and bacterial strains --- p.39 / Chapter 3.2.4 --- Plant and animal materials --- p.40 / Chapter 3.2.5 --- Computer software --- p.40 / Chapter 3.3 --- Purification of LRP --- p.41 / Chapter 3.3.1 --- Purification of LRP from winged bean --- p.41 / Chapter 3.3.1.1 --- Extraction of total protein --- p.41 / Chapter 3.3.1.2 --- Differential pI precipitation --- p.41 / Chapter 3.3.1.3 --- Determination of the pI point of LRP --- p.42 / Chapter 3.3.1.4 --- Native tricine-PAGE and gel elution --- p.42 / Chapter 3.3.2 --- Purification from E. coli --- p.45 / Chapter 3.3.2.1 --- Construction of pET vector expressing recombinant LRP (rLRP) --- p.45 / Chapter 3.3.2.2 --- Expression of rLRP --- p.50 / Chapter 3.3.2.3 --- Purification by gel electrophoresis and gel band elution --- p.50 / Chapter 3.4 --- Anti-serum production --- p.52 / Chapter 3.5 --- Allergy tests --- p.53 / Chapter 3.5.1 --- Pepsin digestion --- p.53 / Chapter 3.5.1.1 --- Determination of optimal concentration of pepsin --- p.53 / Chapter 3.5.1.2 --- Pepsin digestion of allergenic and non-allergenic model proteins --- p.55 / Chapter 3.5.1.3 --- Pepsin digestion of LRP and immunodetection --- p.55 / Chapter 3.5.2 --- Trypsin digestion --- p.56 / Chapter 3.5.2.1 --- Determination of optimal trypsin concentration --- p.56 / Chapter 3.5.2.2 --- Trypsin digestion of allergenic and non-allergenic model proteins --- p.57 / Chapter 3.5.2.3 --- Trypsin digestion of LRP and immuno-detection --- p.57 / Chapter 3.5.3 --- Pepsin and trypsin digestion --- p.58 / Chapter 3.5.3.1 --- Digestions of allergenic model proteins --- p.58 / Chapter 3.5.3.2 --- Digestion of LRP --- p.58 / Chapter 3.5.4 --- IgE binding tests --- p.58 / Chapter 3.6 --- Physiology studies --- p.59 / Chapter 3.6.1 --- Preparation for the studies --- p.59 / Chapter 3.6.1.1 --- Growing winged bean in the field --- p.59 / Chapter 3.6.1.2 --- Growing winged bean in sterile conditions --- p.60 / Chapter 3.6.1.3 --- Production ofLRP-cDNA probe --- p.60 / Chapter 3.6.2 --- Detecting the expression of LRP in winged bean --- p.61 / Chapter 3.6.2.1 --- RNA extraction --- p.61 / Chapter 3.6.2.2 --- RT-PCR and DNA sequencing --- p.62 / Chapter 3.6.2.3 --- RNA electrophoresis and northern blot analysis --- p.63 / Chapter 3.6.2.4 --- Protein extraction --- p.63 / Chapter 3.6.2.5 --- Western blot and immuno-detection --- p.63 / Chapter 3.6.3 --- Expression of LRP in germinating winged bean seeds --- p.64 / Chapter 3.6.3.1 --- Seed germination --- p.64 / Chapter 3.6.3.2 --- Detection of LRP in germinating seeds --- p.64 / Chapter 3.6.4 --- RNase activity test --- p.65 / Chapter 4 --- Results --- p.67 / Chapter 4.1 --- Purification of LRP --- p.67 / Chapter 4.1.1 --- Purification from winged bean --- p.67 / Chapter 4.1.1.1 --- Identification of pI point of LRP --- p.67 / Chapter 4.1.1.2 --- Native tricine PAGE and gel elution --- p.70 / Chapter 4.1.2 --- Purification from E. coli --- p.71 / Chapter 4.1.2.1 --- Construction of pET-LRP vector --- p.71 / Chapter 4.1.2.2 --- Expression of rLRP and gel purification --- p.74 / Chapter 4.2 --- Antiserum production --- p.76 / Chapter 4.3 --- Allergy tests --- p.81 / Chapter 4.3.1 --- Pepsin digestion --- p.81 / Chapter 4.3.2 --- Trypsin digestion --- p.89 / Chapter 4.3.3 --- Pepsin and trypsin digestion --- p.96 / Chapter 4.3.4 --- Human serum IgE binding test --- p.104 / Chapter 4.4 --- Physiological studies --- p.105 / Chapter 4.4.1 --- Samples preparation --- p.105 / Chapter 4.4.2 --- RT-PCR and DNA sequencing --- p.105 / Chapter 4.4.3 --- Expression profile of WBLRP in winged bean somatic organs --- p.108 / Chapter 4.4.4 --- Expression profile ofWBLRP in winged bean flower --- p.111 / Chapter 4.4.5 --- Expression profile ofWBLRP in winged bean maturing seeds --- p.114 / Chapter 4.4.6 --- Expression profile of WBLRP gene in winged bean germinating seeds --- p.117 / Chapter 4.4.7 --- Functional assay of LRP --- p.121 / Chapter 5 --- Discussion --- p.124 / Chapter 5.1 --- LRP purification and antibody production --- p.124 / Chapter 5.2 --- Allergy tests --- p.125 / Chapter 5.3 --- Expression of LRP in WB --- p.131 / Chapter 5.4 --- Functional assay of LRP --- p.134 / Chapter 5.5 --- Hypothesis Testing --- p.135 / Chapter 5.6 --- Future prospective， --- p.136 / Chapter 6 --- Conclusion --- p.138 / Chapter 7 --- References --- p.140