SB2013 L04gr5: Difference between revisions
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<StructureSection load='1l8w' size='400' side='right' scene='' caption='Variable surface antigen (VlsE) (PDB code [[1l8w]])'> | |||
=VlsE= | =VlsE= | ||
[http://en.wikipedia.org/wiki/Lyme_disease Lyme disease] is the most pervasive tick-borne disease in Europe, the United States,and parts of Asia.<ref name="Coutte" /> It is a multistage infection caused by the spirochete [http://en.wikipedia.org/wiki/Borrelia_burgdorferi ''Borrelia burgdorferi'']. Early symptoms include headaches, depression, rash, and fever. If Lyme disease is left untreated, serious complications of the joints, heart and central nervous system can occur. Infected [http://en.wikipedia.org/wiki/Ixodes ''Ixodes'' ticks] in their nymph stage transmit Lyme disease by attaching to humans and other mammals<ref name="Lab Tutor" />. In most cases, the tick must be attached for 36-48 hours before the bacteria can be transmitted.<ref name="CDC" /> | [http://en.wikipedia.org/wiki/Lyme_disease Lyme disease] is the most pervasive tick-borne disease in Europe, the United States,and parts of Asia.<ref name="Coutte" /> It is a multistage infection caused by the spirochete [http://en.wikipedia.org/wiki/Borrelia_burgdorferi ''Borrelia burgdorferi'']. Early symptoms include headaches, depression, rash, and fever. If Lyme disease is left untreated, serious complications of the joints, heart and central nervous system can occur. Infected [http://en.wikipedia.org/wiki/Ixodes ''Ixodes'' ticks] in their nymph stage transmit Lyme disease by attaching to humans and other mammals<ref name="Lab Tutor" />. In most cases, the tick must be attached for 36-48 hours before the bacteria can be transmitted.<ref name="CDC" /> | ||
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Within the variable domain, there are invariable regions that remain unchanged during antigenic variation, and therefore may be targets of an immune response. IR6, the most conserved IR, has been found to be immunodominant. The antigencity of each of the 6 IRs has been studied using peptide-based enzyme linked immunosorbent essays, [http://en.wikipedia.org/wiki/ELISA ELISA].<ref name="Liang" /> | Within the variable domain, there are invariable regions that remain unchanged during antigenic variation, and therefore may be targets of an immune response. IR6, the most conserved IR, has been found to be immunodominant. The antigencity of each of the 6 IRs has been studied using peptide-based enzyme linked immunosorbent essays, [http://en.wikipedia.org/wiki/ELISA ELISA].<ref name="Liang" /> | ||
__TOC__ | __TOC__ | ||
=Structure= | =Structure= | ||
The lipoprotein, VlsE, consists of two invariable domains at the amino and carboxyl termini and a variable domain (Figure 1). When referring to the primary structure of the protein, the variable domain is a cassette region located between the two termini. The variable domain can be further broken down into variable regions (<scene name='SB2013_L04gr5/Variable_regions/3'>VRs</scene>) and invariable regions (<scene name='SB2013_L04gr5/Invariable_regions/2'>IRs</scene>) <ref name="Liang" />. | The lipoprotein, VlsE, consists of two invariable domains at the amino and carboxyl termini and a variable domain (Figure 1). When referring to the primary structure of the protein, the variable domain is a cassette region located between the two termini. The variable domain can be further broken down into variable regions (<scene name='SB2013_L04gr5/Variable_regions/3'>VRs</scene>) and invariable regions (<scene name='SB2013_L04gr5/Invariable_regions/2'>IRs</scene>) <ref name="Liang" />. | ||
When crystallized, VlsE forms a four molecule asymmetric unit with each molecule having slight differences in their conformation. | When crystallized, VlsE forms a four molecule asymmetric unit with each molecule having slight differences in their conformation. Although each molecule in the unit is slightly different, a single molecule of the protein consists of eleven [http://en.wikipedia.org/wiki/Alpha_helices α-helices] and four short [http://en.wikipedia.org/wiki/Beta_strand β-strands]. Helices α1 (aa 306-341), α2 (aa 68-87), α3 (aa 114-139), and α11 (aa 306-341) all form the <scene name='SB2013_L04gr5/Membrane_proximal_region/1'>membrane proximal region</scene> of VlsE, while helices α4 (aa 161-176), α5 (aa 180-185), α6 (aa 195-201), α7 (aa 213-224), α8 (aa 228-239), α9 (aa 255-260) and α10 (aa 277-289) form the primary region of the <scene name='SB2013_L04gr5/Membrane_distal_region/2'>membrane distal region</scene> of the protein. The four short β-strands each consist of 3 amino acids and can also be located in the membrane distal region <ref name="Eicken" />. | ||
Covering the membrane distal part of VlsE are connecting loop regions, which lack secondary structure and have different conformations in each of the molecules. Helices α3 through α10 form the invariable regions and are attached by the connecting loops that are classified as the variable regions. Although VlsE crystallizes into an asymmetrical unit, it appears primarily as monomeric in solution. Because the interface between VlsE molecules in the crystal structure buries approximately 13% of the accessible surface area of the monomers, research suggests that there is a possibility of VlsE existing as a [http://en.wikipedia.org/wiki/Protein_dimer dimer] when in its natural state (Figure 2)<ref name="Eicken" />. <scene name='SB2013_L04gr5/Reset_button/1'>Reset</scene> | Covering the membrane distal part of VlsE are connecting loop regions, which lack secondary structure and have different conformations in each of the molecules. Helices α3 through α10 form the invariable regions and are attached by the connecting loops that are classified as the variable regions. Although VlsE crystallizes into an asymmetrical unit, it appears primarily as monomeric in solution. Because the interface between VlsE molecules in the crystal structure buries approximately 13% of the accessible surface area of the monomers, research suggests that there is a possibility of VlsE existing as a [http://en.wikipedia.org/wiki/Protein_dimer dimer] when in its natural state (Figure 2)<ref name="Eicken" />. <scene name='SB2013_L04gr5/Reset_button/1'>Reset</scene> | ||
[[Image:VlsE 2-d image.jpg]] | [[Image:VlsE 2-d image.jpg|left|500px|thumb]] | ||
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Figure 1 shows the different domains of VlsE using a simplified 2-D representation of the lipoprotein. | |||
Figure 1 shows the different domains of VlsE. | |||
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The variable regions are an important component of the variable domain of VlsE. These regions have relatively high surface exposure, which accounts for approximately 37% of the total surface area of the crystallized protein [http://en.wikipedia.org/wiki/Monomer monomer]. The strategic location of the variable regions has been implicated in protecting the highly conserved invariable regions from antibody binding. These regions, which are almost completely buried within the membrane distal region, are surrounded by the protective loops of the variable regions.<ref name="Eicken" /> | The variable regions are an important component of the variable domain of VlsE. These regions have relatively high surface exposure, which accounts for approximately 37% of the total surface area of the crystallized protein [http://en.wikipedia.org/wiki/Monomer monomer]. The strategic location of the variable regions has been implicated in protecting the highly conserved invariable regions from antibody binding. These regions, which are almost completely buried within the membrane distal region, are surrounded by the protective loops of the variable regions.<ref name="Eicken" /> | ||
The variable regions’ location also supports the potential role that the protein plays in evading host immune responses through [http://en.wikipedia.org/wiki/Gene_conversion gene conversion].<ref name="Zhang and Norris" /> These regions are hypothesized to expose highly variable [http://en.wikipedia.org/wiki/Epitopes epitopes], which would prevent the immune system from recognizing the antigen and subsequently prevent the onset of Lyme disease.<ref name="Eicken" /> This feature starkly contrasts to that of the invariable regions, which does not undergo constant variation and whose genetic composition remains mostly conserved.<ref name="Zhang and Norris" /> | The variable regions’ location also supports the potential role that the protein plays in evading host immune responses through [http://en.wikipedia.org/wiki/Gene_conversion gene conversion].<ref name="Zhang and Norris" /> These regions are hypothesized to expose highly variable [http://en.wikipedia.org/wiki/Epitopes epitopes], which would prevent the immune system from recognizing the antigen and subsequently prevent the onset of Lyme disease.<ref name="Eicken" /> This feature starkly contrasts to that of the invariable regions, which does not undergo constant variation and whose genetic composition remains mostly conserved.<ref name="Zhang and Norris" /> | ||
=Invariable Regions= | =Invariable Regions= | ||
Interspersed in the variable regions of the variable domain are six invariable regions (IR1-IR6). Although located on the membrane distal region of the protein, the IRs are buried within the protein and have little exposure to the surface. The IRs might be further hidden from surface exposure due to the possible dimerization of VlsE, forming a shield at the monomer-monomer interface.<ref name="Eicken" />These six IRs do not undergo changes during antigenic variation and are present in many strains and genospecies of ''B. burgdorferi''<ref name="Zhang and Norris" />. | Interspersed in the variable regions of the variable domain are six invariable regions (IR1-IR6). Although located on the membrane distal region of the protein, the IRs are buried within the protein and have little exposure to the surface. The IRs might be further hidden from surface exposure due to the possible dimerization of VlsE, forming a shield at the monomer-monomer interface.<ref name="Eicken" />These six IRs do not undergo changes during antigenic variation and are present in many strains and genospecies of ''B. burgdorferi''<ref name="Zhang and Norris" />. | ||
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=VlsE as a Diagnostic Tool= | =VlsE as a Diagnostic Tool= | ||
== ''IR 6''== | == ''IR 6''== | ||
IR6 is instrumental in the serodiagnosis of Lyme disease in its early stages. The sensitivity and precision of ELISA was measured based on a 26-mer synthetic peptide (C6) whose sequence matched that of IR6.<ref name="Marangoni" /> Serum samples in nonhuman primates that were introduced to different strains of ''B. burgdorferi'' were assessed for antibody responses. Antibody [http://en.wikipedia.org/wiki/Immunoglobulin_G IgG]was present in high levels for all animals over a 160-week period following infection. Human patients assessed with the C6 ELISA yielded 74% detection sensitivity in the acute disease type, 90% in the convalescent (recovery) phase, 95% in the early disseminated phase, and 100% in late Lyme disease. No antibody responses to peptides resembling the sequences of the remaining IRs were detected in humans or monkeys.<ref name="Liang" /> | IR6 is instrumental in the serodiagnosis of Lyme disease in its early stages. The sensitivity and precision of ELISA was measured based on a 26-mer synthetic peptide (C6) whose sequence matched that of IR6.<ref name="Marangoni" /> Serum samples in nonhuman primates that were introduced to different strains of ''B. burgdorferi'' were assessed for antibody responses. Antibody [http://en.wikipedia.org/wiki/Immunoglobulin_G IgG]was present in high levels for all animals over a 160-week period following infection. Human patients assessed with the C6 ELISA yielded 74% detection sensitivity in the acute disease type, 90% in the convalescent (recovery) phase, 95% in the early disseminated phase, and 100% in late Lyme disease. No antibody responses to peptides resembling the sequences of the remaining IRs were detected in humans or monkeys.<ref name="Liang B" /> | ||
==''N- and C- Terminal Regions''== | ==''N- and C- Terminal Regions''== | ||
In addition to the IR6 region, specific sequences of the N- and C- terminal invariable domains are known to be major B cell epitopes in Lyme disease patients. | In addition to the IR6 region, specific sequences of the N- and C- terminal invariable domains are known to be major B cell epitopes in Lyme disease patients. Although the N and C termini appeared to be flexible when crystallized<ref name="Eicken" />, the crystal structure of VlsE indicates that the two sequences lie adjacent to each other, making them a single target covered by peptides VlsE21 through VlsE31 and VlsE336 through VlsE343.<ref name="Chandra" /> It has been found that antibodies that interact with this membrane-proximal part become more potent in later stages of infection. According to ELISA results, post Lyme disease syndrome patients exhibited much higher antibody activity at the epitopes than fully recovered patients. Therefore, detection of these antibodies are useful in patient follow-ups, especially with those experiencing the later stages of Lyme disease.<ref name="Chandra" /> | ||
=Conclusion= | =Conclusion= | ||
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Though VlsE has been the object of intense scrutiny by researchers, there is still much to be understood. Future experiments may include further studies of the invariable regions, with a specific focus on IR6. IR6 is known to be highly conserved and thus, elicit immunogenic responses in the host once it is exposed. However, the function of IR6 for the survival of the spirochete is unknown. Focusing on how the invariable regions contribute to the virulence of ''B. borgdorferi'' may continue to increase our understanding of the function of VlsE. | Though VlsE has been the object of intense scrutiny by researchers, there is still much to be understood. Future experiments may include further studies of the invariable regions, with a specific focus on IR6. IR6 is known to be highly conserved and thus, elicit immunogenic responses in the host once it is exposed. However, the function of IR6 for the survival of the spirochete is unknown. Focusing on how the invariable regions contribute to the virulence of ''B. borgdorferi'' may continue to increase our understanding of the function of VlsE. | ||
Designing the | Designing the Proteopedia page has greatly increased our understanding of VlsE. Through the process of creating green links to highlight the important functional aspects of VlsE’s structure, the process of relating the structural components of VlsE to its function became more coherent. Researching other articles that studied different aspects of VlsE also allowed us to better comprehend the function of this lipoprotein. In addition, working as a team helped to solidify certain concepts, as members of the group were able to help each other through the process of learning. Creating the Proteopedia webpage project has thus been a useful supplement in enhancing our understanding of VlsE. | ||
</StructureSection> | |||
__NOTOC__ | |||
==See also [[VlsE]]== | |||
=References= | =References= | ||