隨著社會發展, 各種病毒、環境致癌物, 以及不健康食等多種因素導致諸種頑症高發, 並以人類獲得牲免疫缺陷綜合症(艾滋病)和各種腫瘤為代表。從天然產物, 特別是傳統中藥中, 篩選藥用有效成分是治療這類疾病的有效途徑之一。研究發現, 核糖體失活蛋白, 核糖核酸酶, 凝集素, 蛋白酪抑制劑等具有良好的藥用開發前景。本論文著重於從不同植物種于中篩選藥用蛋白, 並對其藥用機制進行研究。 / 是吹研究共純化出六種藥用蛋白。第一, 從苦瓜種子中純化出一種二型核糖體失活蛋白MCL。體外細胞試驗和體內裸鼠試驗顯示MCL 能夠有效抑制鼻咽癌細胞CNE-l 和CNE-2 生長。第二, 苦瓜種于中一個新的核糖核酸酪RNase MC2被分離出來。RNase MC2 通過調控半胱氨酸依賴性細胞死亡蛋白晦(Caspase)信號途徑和絲裂原活化蛋白激酶(MAPKs) 信號途徑誘導乳腺癌MCF-7 細胞凋亡。第三, 從宮粉羊蹄甲種子中提取一種具有抑制腫瘤細胞生長的蛋白酪抑制劑BvvTI 。第四, 從紅花羊蹄甲種子中提取出一與BvvTI 類似的具有抗腫瘤生長的蛋白酶抑制劑BPLTI。 第五，特長秋紫莢豆中存在一個血液凝集素EAPl。EAPL具有制止HIV-l 逆轉錄酶活性, 抗腫瘤, 誘導一氧化氮生成功效。第六, 從另外一種四季豆, 藍虎王, 中提取出一個血液凝集素BTKL。BTKL 通過誘導肝癌HepG2細胞出現DNA 斷裂, 細胞核破壞, 升高線粒體膜通透性, 誘導一氧化氮和細胞因予的表達, 從而導致其凋亡。 / 總之, 上述實驗結果表明, 這六種蛋白具有一定的藥用前景, 可以作為治療艾滋病和不同腫瘤的候造藥物或者候選輔助藥物。進一步體內實驗和臨床實驗評價其療效值得開展。 / Viral pathogens, environmental carcinogens, and unhealthy diets cause severe damage to humans, leading to the acquirement of different stubborn diseases exemplified by AIDS/HIV and neoplasms. Screening of new drugs from natural products, especially from traditional Chinese medicine, provides a promising strategy for these patients. Proteins with potential medicinal applications include ribosome-inactivating proteins (RIPs), ribonucleases, lectins, protease inhibitors and others. The intent of this research proposal is to isolate and characterize proteins with therapeutic potential from plant seeds. / In this project, six medicinal proteins of different origins have been purified by liquid chromatography. One of them is Momordica charantia lectin (MCL), which is a type 11 RIP from the seeds of bitter gourd (M. charantia, BG), with antitumor activity toward human nasopharyngeal carcinoma cells in vitro and in vivo. We have purified a new ribonuclease, named RNase MC2, from BG seeds. It selectively induces apoptosis in breast cancer cells associated with caspase pathways induction and MAPKs activation. Two Kunitz-type trypsin inhibitors, termed BvvTI and BPLTI, have been purified and characterized from the seeds of Bauhinia variegata var. variegata, and B. purpurea L., respectively. EAPL, a lectin with anti-HIV-l reverse transcriptase, antitumor, and nitric oxide (NO) inducing activities was purified from seeds of Phaseolus vulgaris cv. Extralong autumn purple bean. Finally, BTKL is a new P. vulgaris lectin that induced selective toxicity on human liver carcinoma Hep G2 cells. / In conclusion, the above results evince that these proteins are good candidates for the exploration of anti-HIV and/or antitumor drugs or adjuvants. Further research on their efficacies in in vivo as well as clinical trials is warranted. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Fang, Fei. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 162-187). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / ABSTRACT --- p.ii / 中文摘要 (CHINESE ABSTRACT) --- p.iii / ACKNOWLEDGEMENTS --- p.iv / PUBLICATIONS --- p.v / LIST OF ABBREViATIONS --- p.ix / LIST OF FIGURES --- p.x / LIST OF TABLES --- p.xii / Chapter CHAPTER1 --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Prelude --- p.2 / Chapter 1.2 --- Literature review of bitter gourd --- p.4 / Chapter 1.2.1 --- Anti-diabetic property of BG --- p.5 / Chapter 1.2.2 --- Anti-HIV activity of BG --- p.15 / Chapter 1.2.3 --- Antitumor activity of BG --- p.18 / Chapter 1.2.4 --- Looking forward --- p.25 / Chapter 1.2.5 --- Conclusions --- p.29 / Chapter 1.3 --- Research rationale, design, and brief results --- p.35 / Chapter CHAPTER2 --- PURIFICATION AND CHARACTERIZATION OF RIBOSOME INACTIVATING PROTEIN --- p.39 / Chapter 2.1 --- Momordica charantia lectin, a type 11 ribosome inactivating protein, exhibits antitumor activity toward human nasopharyngeal carcinoma cells in vitro and in vivo --- p.40 / Chapter 2.1.1 --- Introduction --- p.40 / Chapter 2.1.2 --- Materials and methods --- p.42 / Chapter 2.1.3 --- Results --- p.46 / Chapter 2.1.4 --- Discussion --- p.59 / Chapter CHAPTER 3 --- PURIFICATION AND CHARACTERIZATION OF RIBONUCLEASE --- p.64 / Chapter 3.1 --- RNase MC2: A new Momordica charantia ribonuclease that selectively induces apoptosis in breast cancer cells associated with MAPKs activation and caspase pathways induction --- p.66 / Chapter 3.1.1 --- Introduction --- p.66 / Chapter 3.1.2 --- Materials and methods --- p.67 / Chapter 3.1.3 --- Results --- p.69 / Chapter 3.1.4 --- Discussion --- p.79 / Chapter CHAPTER 4 --- PURIFICATION AND CHARACTERIZATION OF PROTEASE INHIBITORS --- p.84 / Chapter 4.1 --- Bauhinia variegata var variegata trypsin inhibitor: from isolation to potential medicinal applications --- p.86 / Chapter 4.1.1 --- Introduction --- p.86 / Chapter 4.1.2 --- Materials and methods --- p.86 / Chapter 4.1.3 --- Results --- p.89 / Chapter 4.1.4 --- Discussion --- p.97 / Chapter 4.2 --- A potential human hepatocellular carcinoma inhibitor from Bauhinia purpurea L.seeds: From purification to mechanism exploration --- p.99 / Chapter 4.2.1 --- Introduction --- p.99 / Chapter 4.2.2 --- Materials and methods --- p.100 / Chapter 4.2.3 --- Results --- p.101 / Chapter 4.2.4 --- Discussion --- p.112 / Chapter CHAPTER 5 --- PURIFICATION AND CHARACTERIZATION OF MEDICINAL LECTINS --- p.114 / Chapter 5.1 --- A Lectin with Anti-HIV-l Reverse Transcriptase, Antitumor and Nitric Oxide Inducing Activities from Seeds of Phaseolus vulgaris cv Extra-long Autumn Purple Bean --- p.116 / Chapter 5.1.1 --- Introduction --- p.118 / Chapter 5.1.2 --- Materials and methods --- p.119 / Chapter 5.1.3 --- Results --- p.123 / Chapter 5.1.4 --- Discussion --- p.132 / Chapter 5.2 --- A new Phaseolus vulgaris lectin induces selective toxicity on human liver carcinoma Hep G2 cells --- p.136 / Chapter 5.2.1 --- Introduction --- p.136 / Chapter 5.2.2 --- Materials and methods --- p.137 / Chapter 5.2.3 --- Results --- p.138 / Chapter 5.2.4 --- Discussion --- p.150 / Chapter CHAPTER6 --- CONCLUSION AND FUTURE PERSPECTiVES --- p.154 / Chapter 6.1 --- Conclusions --- p.156 / Chapter 6.2 --- Future perspectives --- p.159 / References --- p.162

Thesis (M.S.)--Brigham Young University. Dept. of Plant and Wildlife Sciences, 2007. / Includes bibliographical references (p. 26-32).

Thesis (M.S.)--Brigham Young University. Dept. of Plant and Animal Sciences, 2006. / Includes bibliographical references (p. 31-37).

Plants accumulate and store proteins in protein storage vacuoles (PSVs) during seed development and maturation. Upon seed germination, these storage proteins are mobilized to provide nutrients for seedling growth. However, little is known about the molecular mechanisms of protein degradation during seed germination and post-germination. Here I test the hypothesis that vacuolar sorting receptor (VSR) proteins play a role in mediating protein degradation in germinating and post-germination seeds. It is demonstrated that both VSR proteins and hydrolytic enzymes are synthesized de novo during mung bean seed germination and post-germination. Immunogold electron microscopy (EM) with VSR antibodies demonstrates that VSRs mainly locate to the peripheral membrane of multivesicular bodies (MVBs), presumably as recycling receptors in Day-1 germinating seeds, but become internalized to the MVB lumen, presumably for degradation at Day-3 post-germination. Chemical cross-linking and immunoprecipitation with VSR antibodies have identified the cysteine protease aleurain as a specific VSR-interacting protein in germinating and post-germination seeds. Further immunogold EM studies demonstrate that VSR and aleurain colocalize to MVBs, as well as PSVs in germinating and post-germination seeds. Thus, MVBs in germinating and post-germination seeds exercise dual functions: as a storage compartment for proteases that are physically separated from PSVs in the mature seed, and as an intermediate compartment for VSR-mediated delivery of proteases from the Golgi apparatus to the PSV for protein degradation during seed germination and post-germination. / Storage proteins synthesized during seed development are transported to PSVs for storage. However, relatively little is known about the mechanisms of storage protein transport. A putative VSR-interacting protein termed S2 was identified as mung bean 8S globulin. Thus, I test the hypothesis that VSR proteins may be involved in storage protein transport to PSVs in developing mung bean seeds. Immunogold EM with 52 (8S globulin) antibody demonstrates that transport of 8S globulin to PSVs is Golgi-mediated, involving dense vesicle (DV) and a novel prevacuolar compartment (PVC). The novel PVC consists of storage protein aggregates and small internal vesicles. Immunogold EM with S2 (8S globulin) antibody demonstrates that MVBs contain 8S globulin at early stage of seed development. Further immunogold EM studies demonstrate that VSR and 8S globulin colocalize to DVs and the novel PVCs. In vitro binding study demonstrates that calcium ion can stabilize interaction between VSRs and 8S globulin. Thus, VSR proteins may mediate storage protein transport to PSVs via a novel PVC. / Wang, Junqi. / "March 2007." / Adviser: Jiang Liwen. / Source: Dissertation Abstracts International, Volume: 69-01, Section: B, page: 0052. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 120-131). / 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.

The occurrence of seed-producing wheat x jointed goatgrass hybrids in infested wheat fields suggests the possibility of gene flow between the two species. This study investigates 'Madsen' wheat x jointed goatgrass F��� and reciprocal backcross derivatives produced in experimental field plantings. Electrophoresis of the high molecular weight (HMW) glutenin seed proteins, chromosome counts, and morphological studies were used to better understand the genetics of these hybrids, and to provide a baseline for evaluating hybrids collected in natural populations. The HMW glutenin profiles are a useful diagnostic tool because the banding patterns, in the 68-120 kDa molecular weight range, are species-unique (three bands for goatgrass, four bands for wheat) and can be used to trace parentage in the hybrid seed on the basis of band contribution. Experimental hybrids show considerable diversity in banding profiles (9 patterns of three to six bands). Diversity in number of different glutenin profiles and number of subunits per seed decreases in more advanced generations (BC��� and BC���S���). Chromosome counts confirm the direction of the crosses and vary as follows: 35 chromosomes for F���; 36 to 57 for BC���; 28 to 49 for BC���; and 33 to 52 for BC���S���. A chromosome number of 28 suggests that jointed goatgrass (2n=4x=28) was the recurrent backcross pollen donor, while numbers closer to 42 and above point to wheat (2n=6x=42) as the pollen donor. Partial female fertility was found in all generations, as well as full self-fertility in BC��� and BC���S��� plants. Analysis of the HMW glutenin profiles of the progeny seeds verifies that hybridization can go in either direction, with most banding patterns similar to either jointed goatgrass or wheat. The resulting potential for gene flow from wheat to jointed goatgrass calls for continued study of these hybrid derivatives. / Graduation date: 2001

Chickpea (Cicer arietinum L.) seed is a potential source of protein ingredients with desirable nutritional and functional properties. Knowledge of molecular characteristics of a food protein is essential before a protein can gain widespread use as a food ingredient. The objectives of this study were to prepare chickpea proteins using different extraction methods and precipitation methods and to investigate molecular characteristics using polyacrylamide gel electrophoresis (PAGE; Native and SDS), reversed phase high performance liquid chromatography (RP-HPLC) and electrospray ionization mass spectrometry (ESI-MS) techniques. Proteins of ground chickpea seed were extracted with sodium hydroxide (NaOH) and with citric acid solutions and precipitated with addition of acid and by cryoprecipitation. The protein contents of the protein preparation ranged from 49% to 97%. The microstructures of chickpea protein isolates examined by scanning electron microscope (SEM) revealed the presence of starch grains in the cryoprecipitates from citric acid extraction but not in isoelectric precipitates. The globulins (legumins and vicilins), glutelins, and albumins from both citric acid and NaOH isolates were characterized by Native-PAGE. The cryoprecipitates contained mainly the globulin-rich proteins. With SDS-PAGE characterization, protein subunits were identified as follows: (i) legumin subunits: MW 40, 39, 26, 23, and 22 kDa, (ii) vicilin subunits: MW 50, 37, 33, 19, and 15 kDa, (iii) glutelin subunits: 58, 55, and 54 kDa, and (iv) albumin subunits: 10 kDa. Separation of fractions of isolated chickpea proteins by RP-HPLC showed that early eluting fractions (Rt 20-30 min) consisted of subunits of MW 6.5-31 kDa (SDS-PAGE). At elution time 30-36 min, the fractions obtained were composed mainly of mixtures of legumin and vicilin subunits (MW 14-45 kDa). The major subunits of chickpea protein fractions from both cryoprecipitates and isoelectric precipitates are legumin basic subunit (MW∼23 kDa) and vicilin-rich proteins (MW∼19, 17, 15 kDa). ESI-MS analysis of fractions separated by RP-HPLC showed MW ranging between 5.1 and 53.5 kDa. The subunits of MW 35366, 27626, 22864, 20531, 16092, and 15626 Da of fractions from ESI-MS corresponded to MW 35.3, 28.0, 24.1, 20.5, 16.1, and 15.3 kDa identified in SDS-PAGE. These fractions were identified as legumin-rich and vicilin-rich proteins.

No description available.

by Foo Wai Ting, Rita. / Thesis submitted in: August 1998. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 89-100). / Abstract also in Chinese. / Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Proximate Composition --- p.4 / Chapter 1.2 --- Amino Acid Composition --- p.6 / Chapter 1.3 --- Antinutrients --- p.11 / Chapter 1.3.1 --- Trypsin Inhibitors --- p.12 / Chapter 1.3.2 --- Phytate --- p.13 / Chapter 1.3.3 --- Tannins --- p.14 / Chapter 1.3.4 --- Lectins --- p.15 / Chapter 1.4 --- Two Dimensional Polyacrylamide Gel Electrophoresis --- p.17 / Chapter 1.5 --- Protein Digestibility --- p.19 / Chapter 1.6 --- Protein Quality --- p.22 / Chapter 1.7 --- Hypocholesterolemic Effects --- p.24 / Chapter 2 --- Materials and Methods --- p.36 / Chapter 2.1 --- Plant Material --- p.36 / Chapter 2.2 --- Sample preparation --- p.36 / Chapter 2.3 --- Proximate composition --- p.38 / Chapter 2.3.1 --- Protein --- p.38 / Chapter 2.3.2 --- Fat --- p.38 / Chapter 2.3.3 --- Carbohydrate --- p.38 / Chapter 2.3.4 --- Fiber --- p.38 / Chapter 2.3.5 --- Mineral --- p.39 / Chapter 2.3.6 --- Moisture --- p.39 / Chapter 2.4 --- Amino acid composition --- p.40 / Chapter 2.5 --- Antinutrients --- p.41 / Chapter 2.5.1 --- Trypsin inhibitors --- p.41 / Chapter 2.5.2 --- Tannins --- p.42 / Chapter 2.5.3 --- Phytate --- p.43 / Chapter 2.5.4 --- Lectins --- p.43 / Chapter 2.6 --- Two dimensional polyacrylamide gel electrophoresis --- p.45 / Chapter 2.6.1 --- Protein extraction --- p.45 / Chapter 2.6.2 --- IEF gel --- p.45 / Chapter 2.6.3 --- SDS gel --- p.46 / Chapter 2.7 --- Protein digestibility --- p.48 / Chapter 2.7.1 --- In vitro Protein digestibility --- p.48 / Chapter 2.7.2 --- True Protein digestibility --- p.49 / Chapter 2.8 --- Protein quality --- p.51 / Chapter 2.9 --- Hypocholesterolemic effects --- p.52 / Chapter 2.10 --- Statistical analysis --- p.55 / Chapter 3 --- Results --- p.56 / Chapter 3.1 --- Proximate composition --- p.56 / Chapter 3.2 --- Amino acid composition --- p.56 / Chapter 3.3 --- Antinutrients --- p.56 / Chapter 3.4 --- Two dimensional polyacrylamide gel electrophoresis --- p.60 / Chapter 3.5 --- Protein digestibility --- p.60 / Chapter 3.6 --- Protein quality --- p.60 / Chapter 3.7 --- Hypocholesterolemic effects --- p.62 / Chapter 3.7.1 --- Growth rate against day --- p.62 / Chapter 3.7.2 --- Health indexes --- p.64 / Chapter 3.7.3 --- Cholesterol content --- p.64 / Chapter 4 --- Discussion --- p.67 / Chapter 4.1 --- Proximate composition --- p.67 / Chapter 4.2 --- Amino acid composition --- p.70 / Chapter 4.3 --- Antinutrients --- p.74 / Chapter 4.4 --- Two dimensional polyacrylamide gel electrophoresis --- p.77 / Chapter 4.5 --- Protein digestibility --- p.79 / Chapter 4.6 --- Protein quality --- p.81 / Chapter 4.7 --- Hypocholesterolemic effects --- p.82 / Chapter 5 --- Conclusion --- p.88 / References --- p.89

by Yubing Li. / "September 2003." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (p. 173-190). / 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. / Abstracts in English and Chinese.

Thesis (Ph. D.)--University of Missouri-Columbia, 1997. / Typescript. Vita. Includes bibliographical references (leaves 456-600). Also available on the Internet.