Biochemistry Commons^™

Structural Studies Of Recombinant Human Β-Casein Proteins, Yilin Hu

Loma Linda University Electronic Theses, Dissertations & Projects

β-Casein is the major component (~80%) of the human casein system. Interacting with calcium and phosphates through its phosphorylated N-terminus, β-casein participates in the formation of micelles which carry these otherwise insoluble nutrients indispensable for the growth and development of newborns. Human β-casein, or fragments thereof, are also found to have biological effects such as enhancement of calcium absorption. So far, knowledge of micelle formation is limited and structural studies of β-casein, especially its N-terminal portion will provide a better understanding of the structure-function relationship of milk micelles.

To elucidate the role that the N-terminus of the protein plays in …

A Study On The Effects Of The N-Terminal Amino Acid Sequence On The Activation Of Human T-Cell Leukemia Virus Type 1 Protease, Hidayah Muhammad Kendall

Chemistry & Biochemistry Theses & Dissertations

Human T-cell leukemia virus type 1 (HTL V-1) is dependent upon the enzymatic activity of its protease for maturation. Maturation of the protease is facilitated by cleavage of specific amino acid residues, followed by dimerization. The effects of the amino acid sequence located N-terminally to the cleavage site on the ability of the protease to become active were the focus of the current study. These amino acid sequences were contributed by the plasmid vector into which the protease gene was inserted.

Surface probability analyses (SPAs) of the vectors, as well as for native sequences which produce the mature protease and …

Regulation Of Connexin43 By Phosphorylation, Maithili M. Shah

Loma Linda University Electronic Theses, Dissertations & Projects

Gap junctions play a crucial role in embryogenesis, especially of the heart where connexin43 is thought to be important for its formation as well as synchronized contraction. Several reports have shown that connexin43 protein is phosphorylated on serine residues in vitro and in vivo.

Studies performed in our laboratory have demonstrated that cell-to-cell communication in cells expressing connexin43 can be controlled rapidly and reversibly by microinjection of active protein kinases or phosphatases that target serine or threonine residues. Phosphorylation by cAMP-dependent protein kinase (PKA) seems to favor channel opening, whereas the effect of protein kinase C (PKC) on channel gating …

Nitrate Reductase Activity, Nitrate Uptake And Iodine Speciation In The Marine Environment, Chin-Chang Hung

OES Theses and Dissertations

The present method for the determination of new production (NP) by measuring the uptake of added ¹⁵NO₃^– suffers from a number of limitations. In an attempt to improve this situation, this research examined the possibility of estimating ¹⁵NO₃– uptake by measuring the activity of nitrate reductase (NRA). In addition, because it has long been suspected that the biological reduction of IO₃^– to I^– may be mediated by nitrate reductase (NR), this research investigated the ability of NR to catalyze the reduction of iodate to iodide.

An improved method for the determination …

Generation Of Human Chorionic Gonadotropin Beta Subunit Specific Monoclonal Antibody Using A Synthetic Peptide, Zhe Huang

Chemistry & Biochemistry Theses & Dissertations

Human chorionic gonadotropin (hCG) is well known for early immunological and bioassay tests for early detection of pregnancy. Since the hormone specific beta subunit confers unique biological and immunological specificity to hCG, immunochemical measurement of hCG has achieved greater specificity and sensitivity in recent years by using either the beta subunit as an immunogen or a partially reduced beta subunit polypeptide chain. However, antiserum raised against the entire hCG beta subunit usually cross-reacts to some extent with human luteinizing hormone (hLH). hCG has been found to be present in the extract of tumors and minute quantities of hCG are synthesized …

Regulation Of Synthesis And Actions Of Bone Growth Factors, Rashmi Malpe

Loma Linda University Electronic Theses, Dissertations & Projects

Among the many bone growth factors, this study focused on the IGF and BMP systems because of some of their unique functions, namely, IGFs function both as local and systemic agents and BMPs are capable of inducing de novo bone formation at non-skeletal sites. There is varied osteoblast differentiation in and load bearing on the skeletal system which led to the first study whose hypothesis was that these skeletal site dependent differences are associated with site-specific differences in IGF system components' production. Results from their quantitation in the CM of normal HBC derived from calvaria, mandibles, ribs, vertebrae, and marrow …

Oligosaccharides From Cell Surface Glycoproteins Of Thermoplasma Acidophilum, An Archae, Gitanjali Srivastava

Honors Theses

No abstract provided.

Evidence That Cys230 And Lys238 Of Biotin Carboxylase Do Not Act As An Acid-Base Pair To Deprotonate Biotin, Robert Brett Lloyd

Honors Theses

No abstract provided.

Production Of Copper-Complexing Ligands In Response To Elevated Concentrations Of Copper In Marine Synechococcus Spp., Arunsri Chatkaeomorakot Brown

Theses and Dissertations in Biomedical Sciences

Synechococcus spp. (marine cyanobacteria) are extremely sensitive to copper toxicity and can produce high-affinity ligands of unknown structure which form complexes with free cupric ion. These ligands may contribute to the biological control of the levels of free cupric ions in surface seawater. Synechococcus spp. are known to produce metallothioneins (MT) in response to cadmium and zinc stress. In the present study the hypothesis that marine Synechococcus produce MT in response to copper was tested. Three marine Synechococcus spp. i.e., PCC 7001, PCC 73109, and PCC 7003, were exposed to different concentrations of CUSO 4 for various time periods. Size …

Fabrication Of Protein Arrays Using Electron Beam Lithography, Duy-Bao Phan Dinh

Theses, Dissertations and Capstones

Very few electron beam based methods for producing high-resolution patterns of surface bound proteins on glass have been developed.

There are only two types of end groups at the surface of glass, they are silanol and siloxane groups. An estersil layer can be formed on the surface of glass by treatment with hot alcohol creating a hydrophobic layer. The reaction is depicted as follows.

-Si-OH + HOR -SiO-R +H2O R = alkyl chain

The proposal that protein arrays could be fabricated by patterned electron bombardment using estersils as a negative resist was tested in this research. The hydrophobic estersil layer …

Preparation Of N-(Haloaryl) Succinimides, Okey Noe Ii

Theses, Dissertations and Capstones

N-(3,5-Dichloro-4-fluorophenyl)succinimide (NDCFPS) and N-(3,5-dichloro-4- pyridyl)succinimide (NDPyS) were synthesized using procedures employed previously for N-(3,5-dichIorophenyl)succinimide (NDPS). However, NDPyS was not formed cleanly: extraction from by-products was necessary and purification by column chromatograpy was required. Several potential alternative routes to NDPyS were investigated without success.

Both compounds were administered to male Fischer 344 rats to assess nephrotoxic potential. NDCFPS, contrary to expectations, was non-nephrotoxic. Preliminary results indicate that NDPyS is also non-nephrotoxic.

Environmental Stress In Hard Coral: Evaluating Lipid As An Indicator Of Sublethal Stress On Short Time Scales, David Harold Niebuhr

Dissertations, Theses, and Masters Projects

Lipid quality was evaluated in Montastrea spp. under sediment- and heat-stressed conditions to evaluate lipid ratio as an indicator of sub-lethal stress on short time scales. The ratio of storage lipid (wax ester + triacylglyceride) to structural lipid (sterol esters + phospholipid) decreased significantly (0.25 to 0.14, p < 0.01) after experimental sedimentation. FAME analysis of colonies exposed to experimental sedimentation showed a reduction of the algal, 18:3(n-6) and 18:4(n-3), polyunsaturated fatty acid (PUFA) in the polar lipid fraction. This loss of PUFA suggests a loss of algal membrane in sediment-stressed colonies. Lipid quality was similarly measured in M. faveolata over a 10-day period. Mean (n = 20) ratio of storage to structural lipid in M. faveolata dropped from 2.43 to a level of 0.98 immediately following a natural sedimentation event before recovering to levels of 1.4 and 2.9 on post-storm days 2 and 4, respectively. Colonies of M. annularis subject to heat-stress (35??C) exhibited no significant change in storage lipid ratio, while levels of Free Fatty Acids increased significantly from 0.012 (n = 22) to 0.156 mg lipid/g dry tissue (n = 22)(p < 0.05). FAME analysis of tissue lipids extracted from the heat-stressed colonies showed changes in the polar fraction, with significant decreases in the 18:3(n-6), 18:3(n-3), 18:4(n-3), 20:4(n-6) and 20:5(n-3) (p < 0.05) PUFA and subsequent significant increases in the saturated fatty acids, 16:0 and 18:0 (p < 0.05). These changes in lipid quantity and quality indicate possible oxidation and preferential digestion of zooxanthellar membranes. Stress experiments were repeated in M. annularis using VacutainerRTM blood collection tubes to collect micro-tissue samples without destroying skeleton of the sample colonies. A significant decrease in storage: structural lipid ratio after sedimentation was also detected using the micro-tissue technique. This study indicates that the relative abundance of lipid subclass components can indicate sub-lethal environmental stress, on short time scales, in M. annularis and M. faveolata. Furthermore, micro-tissue collection techniques permit repeated monitoring coral colonies to assess the manifestation of stress from first detection of impact at the cellular level to changes in community to changes in community structure detectable over longer time scales.

The Atomic Force Microscope In The Elucidation Of Leukocyte Abnormality, Derbrenn O'Connor

Theses

The aim of this project was to investigate the novel use of the atomic force microscope (AFM) as a technique in the elucidation of leukocyte abnormality. To this end, comparative AFM imaging and elasticity studies were performed on normal and myeloid leukaemic leukocytes.

Initially, an evaluation of techniques for the isolation of specific leukocyte (e.g. lymphocytes, granulocytes, etc.) populations from whole blood or leukocyte concentrates, was carried out. It was found that a silanised glass surface failed to select and immobilise a leukocyte population. However, centrifugation of whole blood in an Isopaque-ficoll gradient solution yielded a viable mixed leukocyte population …

Investigating The Allosteric Behavior Of Malate Dehydrogenase From Escherichia Coli, Eman M. Ghanem

Masters Theses

Regulatory mechanisms of malate dehydrogenase from E.coli (eMDH) involving NADH as an allosteric effector were investigated. The reaction was studied in both directions: malate oxidation and oxaloacetate reduction. When malate was the variable substrate, a plot of rate against substrate concentration was sigmoidal in the presence of 0.065 mM NADH, which indicates the presence of an allosteric site for NADH on the enzyme. Binding of NADH at the allosteric site causes conformational changes in the active site and, thereby, changes the catalytic activity of the enzyme. An increase in K_m value, from 1.3 to 3.9 mM malate was observed, …