Β-Galactosidase: Protein Analysis

The best one was the twitcher mouse model which occurring human krabbe disease that is caused by a mutation in galactocerebrosidase gene. Mutation analysis of the human GALC gene was facilitated by the cloning and sequencing of GALC cDNA (5). Mutation analysis of the human GALC gene was facilitated by the cloning and sequencing of GALC cDNA . This allowed DNA obtained from Krabbe affected people to be sequenced and analysed against the normal GALC gene. To date there have been over forty mutations identified that cause the galactosylceramidase deficiency of Krabbe disease .The most common mutation in the European population is a 30kb deletion which is associated with a C to T transversion at cDNA position 502. The large 30kb deletion affects the production of galactosylceramidase since it removes a significant portion of the enzyme coding region. This results in the cancellation of 5 amino acids and the insertion of 2 amino acids which impacts on the quaternary structure of the

What is interesting about the disease is that it is the mutation in a single gene called GBA, in which prevents the production of an enzyme called Glucocerebroside (1). In our BIS 102 class, we have discussed the importance of enzymes since enzymes help speed up chemical reactions, especially in our body since enzymes are vital for our bodies to function. Without enzymes, some reactions may take years even hundreds years to react as with enzymes it may take a fraction of a second. Glucocerebrosidase is need to break down a type of lipid that is found in cell membranes called glucocerebroside (2). Glucocerebroside plays a big role in cell membrane since it links to many important functions such as cell to cell interaction, and regulation within the membrane. However, the liver and spleen tend to over produce glucocerebroside, and needs to be to be reduced into adequate amount to have a normal function, in which the enzyme glucocerebrosidase to degrade some of the lipid away. Gaucher disease prevents with the production of the enzyme, and the lipid will accumulate to a point in which affects the functions throughout

It is evident that there are many significant biological syntheses that take place in this world. This includes the synthesis of bilirubin (BR) from biliverdin (BV) with biliverdin IX-beta reductase (BVR). BV is one of three products produced from heme being catalyzed by heme oxygenase. Now, BVR, as seen in Figure 1, is known as an enzyme that catalyzes the reduction of BV’s double bond to a single bond on the second and third pyrrole ring. In other words, BVR reduces BV, which is hydrophilic, using NADPH and yields the product bilirubin, which is hydrophobic (1). The schematic for this mechanism is shown below…

Introduction: The goal of this experiment was to practice using the FirstGlance in Jmol molecular visualization to examine key structural features of proteins. This work is important because protein structure can be related to function, multiple-sequence alignments and evolutionary preservation, and designing drug. FirstGlance in Jmol makes it fairly easy to perceive structure-function relationships in the protein you chose. Using FirstGlance, it is easy to visualize and distinguish chains, and disulfide bonds are obvious. Alpha helices and beta strands are evident due to the "cartoon" secondary structural schematic.

It was the purpose of this experiment to successfully extract the enzyme, -galactosidase from the mutated strain of E. coli sample via a multi-step protein purification process that targeted the enzymes properties of solubility, size and differences in charge. Additionally, the goal of purification was to ensure maximum enzyme production and purity at minimal loss to the enzymatic

HS that contains -GlcA-GlcNS6S-, -GlcA2S-GlcNS- and -GlcA-GlcNS3S- is susceptible for heparanase cleavage17. Heparanase cleaves the bond between non-sulfated glucuronic acid and glucosamine carrying an N-sulfo or both N-sulfo and O-sulfo groups. In addition, heparanase recognizes glucosamine residues that contain N-sulfo and O-sulfo groups at +1 residue of the cleavage site. If the O-sulfation is not present, heparanase recognizes GluA2S located at +/-2 residues of the cleavage site17. In addition, heparanase has two cleavage modes; consecutive cleavage and gapped cleavage. The first step in both modes is similar in which heparanase cleaves the nonreducing trisaccharide end16. In the second step of consecutive mode, the linkage between the fifth and sixth residues is cleaved. On the other hand, the linkage between the seventh and ninth residues is cleaved in the gapped mode16.

In this experiment, the results identify the different temperatures that the lactase enzymes were most effective and ineffective at. At 0°, 1% of glucose was present in the milk meaning that the lacteeze enzymes were effective at the temperature but the reaction time of the enzymes were slower. At 40°, the milk contained the maximum amount of glucose at 2% or more. This indicates that at this temperature, the lacteeze enzymes work the most effective and they have the most amount of collisions which increases the reaction time and breaks down the lactose quicker. At 100°, there was no glucose present in the milk. This meant that all the lactase enzymes were denatured in the hot temperature and none of the lactose sugars were broken down.

The GM2 activator protein also assists in the transport of the GM2 ganglioside to the lysosome, allowing for further degradation by β-hexosaminidase A. Lysosomes are organelles found in eukaryotic cells that contain important enzymes, such as β-hexosaminidase A, that help to breakdown various substances. Upon reaching the lysosome, the β-subunit of β-hexosaminidase A binds to the GM2 activator protein. The β-subunit of β-hexosaminidase A further activates the α-subunit of β-hexosaminidase A. As a final step in degradation, the α-subunit of β-hexosaminidase A cleaves the terminal N-acetylgalactosamine group from the GM2 ganglioside. This cleavage allows for the GM2 ganglioside to be fully digested by the

Polyphen2.0 (Polymorphism Phenotyping) a multi-sequence alignment server predicts the functional impact of an amino acid substitution. The prediction is a straightforward empirical rule which is automatically applied to the structural, sequence, and phylogenetic information, and readily characterizes the amino acid substitution (Ramensky et al. 2002). To determine the effect of variants on the protein secondary structure, inter chain contents, functional sites, and binding sites, Polyphen2.0 utilizes PDB (Protein Data Bank), DSSP (Dictionary of secondary structures in protein), and three-dimensional structure databases (Ramensky et al. 2002). We submitted the query in the form of the protein sequence with the mutational position of two amino

Beta-Galactosidase is a bacterial protein that is coded for the Beta-Galactosidase open reading frame, which is part of the greater overall Lac-Z gene. Beta-Galactosidase cleaves the sugar lactose for metabolic processes. If the homogenic substrate known as X-gal is added to the agar plate, the x-gal is hydrolyzed to form 5- bromo-4-chloro-indoxyl, which spontaneously dimerizes to produce an insoluble blue pigment called 5, 5’-dibromo- 4, 4’-dichloro-indigo and that produces a blue color (Padmananbhan, 2011). This blue color is a visual phenotypic indicator showing whether the beta-galactosidase open reading frame is being transcribed or not. Restriction enzymes are able to recognize specific palindromic sequences in DNA base pairs which are

It is present in oligomer as nonself-recognizing lectin from the hemolymph plasma and it has propeller-like arrangement. This structure gives the protein flexibility which is very important for the specific interactions tachylectin-5A makes with the pathogen associated

The HMG CoA reductase inhibitor’s ring structure would bind to HMGR enzyme. The ring structures involved in HMG CoA reductase inhibition, which are clinically useful, include partially reduced pyrrole in atorvastatin.

Bettelheim, Brown, Campbell and Farrell assert that polypeptide chains do not extend in straight lines but rather they fold in various ways and give rise to a large number of three-dimensional structures (594). This folding or conformation of amino acids in the localized regions of the polypeptide chains defines the secondary structure of proteins. The main force responsible for the secondary structure is the non-covalent

The initial domain architecture was generated based on the known pilE protein in Neisseria gonorrhoeae (control sequence) and consisted of two domains, the N-terminal domain and the pilA domain. Using the metaserver InterPro, the N-terminal domain was predicted to span amino acid residues 2-29, while the pilA domain was predicted to span residues 36-161 (Finn et al., 2017). Although there seemed to be a somewhat high correspondence between the secondary structure prediction analysis and the domain architecture

Just as members in the same family often resemble each other, similarly structured molecules are often grouped in the same category. There are several methods of comparing amino acid sequencing and determining how closely they are related. A simple example is the sequence-comparison method. Sequence databases are searched for a specific amino acid sequence of unknown characteristics. A set of molecules that are similar in structure can give insight into the properties of the unknown sequence of amino acids.