Role Of Sacsin In Lysosomal Transport

If the solution in the left beaker contained both urea and albumin, which membrane(s) could you choose to selectively remove the urea from the solution in the left beaker? How would you carry out this experiment?

Explain the effect that increasing the Na+ Cl- concentration has on osmotic pressure and why it has this effect. How well did the results compare with your prediction?

Cells are a highly complex entity and is both structurally and functionally the basic unit of all living things. Cells are an important and vital part in order to sustain life. Cells are highly diverse and their specific roles are dependent on shape and internal composition. Cells are what carry out functions to promote life, including: reproduction, nutrient digestion, and metabolism. The cell has many components including the nucleus, cytoplasm, many organelles, and a plasma membrane. The nucleus contains all of the genetic material and genes, which are found in the DNA. The nucleus is the control center of the cell as it is key for cell reproduction. The nucleus is surrounded in a membrane called the nuclear envelope for protection and regulation of materials in and out of the nucleus. Cytoplasm is what all of the cell contents is referred to outside of the nucleus. This includes all the high specific metabolic machinery called organelles. A list of organelles include: Ribosomes, endoplasmic reticulum, Golgi apparatus, lysosomes, peroxisomes, mitochondria, and centrioles. Ribosomes are free floating spherical bodies of RNA and proteins which are the site for protein synthesis. Endoplasmic reticulum is broken into two structures, the rough and smooth endoplasmic reticulum. Rough ER provides area for storage and transport of proteins

The purpose of these experiments is to examine the driving force behind the movement of substances across a selective or semiperpeable plasma membrane. Experiment simulations examine substances that move passively through a semipermeable membrane, and those that require active transport. Those that move passively through the membrane will do so in these simulations by facilitated diffusion and filtration. The plasma membrane’s structure is composed in such a way that it can discriminate as to which substances can pass into the cell. This enables nutrients to enter the cell, while keeping unwanted substances out. Active

Lysosomes are membranous sacs of enzymes that are typically are involved in the digestion of cellular macromolecules. They are made by both the endoplasmic reticulum and the Golgi complex. Lysosomes have various roles, including the recycling of the cell's organic material and in the intracellular digestion of macromolecules. The digestion of macromolecules is carried out through an intense concentration of enzymes. To be specific, lysosomes contain close to 50 different degrading enzymes that are capable of hydrolyzing proteins, DNA, RNA, polysaccharides, and lipids within the cell.1 All of lysosomal enzymes are acid hydrolases,

The body has two faces, the cis face which fuses with incoming transport vesicles, and the trans face which excretes the secretory vesicles. The cis face fuses with vesicles coming from the ER effectively from many directions due to its convex shape, whereas the concave trans face can direct the secretory vesicles to their destination. When fusing with the cis face, the transport vesicles release their proteins to be absorbed for modification. Each cisternal layer of the Golgi body holds different enzymes which each modify the passing proteins in separate ways. Between the layers the proteins are moved through the gaps by small vesicles. When a protein has been modified correctly, it leaves the Golgi body via secretory vesicles which then carry the modified proteins to the cell membrane or another organelle. The proteins that are transported to the cell membrane are either excreted from the cell, or absorbed into the membrane to aid with its function. Some of the secretory vesicles which hold hydrolytic enzymes stay within the cytoplasm and function as lysosomes.

sHSPs are a ubiquitous class of chaperones found across all kingdoms of life. sHSP range in size from 12-42 kilo Daltons in large oligomers of 12 to >32 subunits and the structure is homologous across all species. The sHSP monomer consists of three domains: a disordered N-terminal arm, a beta-sandwich α-crystallin domain, and a flexible C-terminal extension. The N-terminal domain is the most variable region with little conservation between species. Experimental evidence also suggests N-terminal involvement in substrate binding and protection. The α-crystallin domain is the most highly conserved region and adopts a β-sandwich conformation composed of 7 to 8 anti-parallel β-strands (Basha et al, 2012). The C-terminus contains an I-X-I motif, which helps to satblizie the oligomeric form of the sHSP (Basha et al, 2012).

Figure 1 Structure of G-protein-coupled receptor, showing alpha-helical proteins attached to the three subunits of the G-protein.

Furthermroe, due to the expression if all of these cosntructs ina abacterial system, post translational modifications are lost which may have an effect on the these constructs three dimensional structre. Expressing these constructs in in a mammalian system such as in human neurons would allow for the study of the effect of post translational modification on influencing JIP3 interaction.

To study the effects of hypotonic, hypertonic and isotonic solutions on plant and animal cells.

Co-immunoprecipitation studies were performed to check the hypothesis that binding of SBP2 to SECIS resulted in recruitment of eEFsec-tRNA. We

Methods used in this experiment include use of plasmids, imaging techniques, use of HEK cells, and voltage/current clamps, and finally an SNRC calculation. Plasmids are used because they are easy to manipulate and integrate into plasma membranes1. This allows an efficient and quick insertion of desired substances into the desired protein like GFP into ASAP1. In this experiment GFP was introduced into the S3 S4 loop where

This particular protein of interest is made up of 524 amino acids. When further analyzing this protein, the Jpred results displayed in Figure 2 give a further understanding of the secondary structure of this protein. It is apparent that the secondary structure of alkaline phosphatase, tissue non-specific isozyme isoform 1-precursor contains primarily alpha helices, yet also contains a few beta sheets. The figure displays the alpha helices by long, red tube like figures and the beta sheets as green arrows. Because this figure contains many more long red tubes than green arrows, it is apparent that the secondary structure contains mostly alpha helices. What results in the secondary structure of a protein is the regular pattern of hydrogen bonding between the N-H from the peptide and the C=O groups of the amino acids. These two structures that result in the hydrogen bonding are found relatively close to one another within the linear sequence. Within this regular pattern of hydrogen bonding is where the alpha helices and beta sheets are found. What primarily makes up the secondary structure of our studied protein, alkaline phosphatase, tissue non-specific isozyme isoform 1-precursor, are alpha helices. Alpha helices are rod-like structures that are stabilized as a result of the hydrogen bonds between the N-H and the C=O groups found among the main

The endoplasmic reticulum (ER) is an essential organelle that is a major place for the biogenesis of cellular components including proteins, lipids, and carbohydrates and internal calcium storage. ER is primarily responsible for protein translocation, protein folding and protein post modification. Proper folding of protein in the ER is accomplished with the aid of ER resident proteins or enzymes such as chaperones. Binding of chaperones to

To find the critical binding region of sacsin which interacts with JIP3, six well-characterized N-terminally GST-tagged sacsin deletion constructs, which include the three SIRPTs, DNAJ-HEPN, UBE3A binding site, and UBL domains (Fig.1), were expressed in a bacterial system and used in pulldown assays to test for JIP3 binding. These domains