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| =='''3-Dimensional Model of Karyopherin-β2'''== | | ==Karyopherin Beta 2 :)== |
| Students: Mary Acheampong, Rosieva Ashong-Katai, Kavita Bhikhi, Daviana Dueño, Bobby Glover, Lachoy Harris, Alafia Henry, Randol Mata, Hillary Ramirez and Marisa VanBrakle, Hostos-Lincoln Academy.
| | <Structure load='4OO6' size='350' frame='true' align='right' caption='Insert caption here' scene='Insert optional scene name here' /> |
| | Karyopherinβ is a group of proteins that is composed of both importins and exportins. Importins are proteins that carry cargos into the nucleus while exportins serve the opposite function. As of today, twenty different Kapβs have been identified. Each of these Kapβs is capable of recognizing and transporting a specific group of cargos. In order to bind to its cargo a Kapβ has to recognize a Nuclear Localization or Export Signal (NLS or NES). |
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| Teacher: Allison Granberry, Hostos-Lincoln Academy
| | Karyopherin beta 2 (kapβ2) is an importin that transports various cargo protein into the nucleus through interactions with nucleoporins, which are proteins of the nucleopore complex (NPC). One might overlook the significance of this protein but actually plays a crucial role in the human body by mediating transport of RNA- binding proteins involved in transcription, RNA processing, RNA transport and translation. The structure of KAPβ is composed of <scene name='37/372723/4oo6_rainbow_heat_repeats/1'>20 antiparallel helices </scene>called [https://en.wikipedia.org/wiki/HEAT_repeat_domain HEAT]repeats. These HEAT repeats contribute to kapβ2’s large superhelical shape. That form two arches:one at the <scene name='37/372723/4oo6_ran_binding_site/2'>N-Terminal</scene> and the other at the |
| | <scene name='37/372723/4oo6_substrate_binding_side/2'>C-Terminal</scene> Through recognition of a nuclear localization signal (NLS) located on its cargo, kapβ2 binds to its cargo via its C-terminal arch. Release of the cargo is mediated by RanGTP, which once bound, modifies the shape of kapβ2. Release of the cargo is mediated once contact occurs between RanGTP and N-terminal arch of Kapβ2. |
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| Mentors: Michael Rout, Natalia Ketaren
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| ==='''Introduction'''===
| | '''How Does Kapβ2 Recognizes Cargo In the Cytoplasm''' |
| | :The NLS located on Kapβ2 cargos are named PY-NLS and they bind to the C-terminal arch of Kapβ2. The electrostatic potential of the internal surface of Kapβ2 superhelix at the C-terminal arch is negative |
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| | Recognition the PY-NLS by Kapβ2 follows a certain guidelines: |
| | #PY-NLS, when not bound to Kapβ2, lacks a secondary structure. |
| | #PY-NLS hass an overall <scene name='37/372723/Positive_interaction_nls_only/3'>positive charge</scene> allowing for electrostatic compatibility with Kapβ2. |
| | #General sequence for the PY-NLS is either a hydrophobic or base at the motif at N-terminus and R-X2-5- P-Y motif at the C-terminus |
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| Karyopherin beta 2 (Kapβ2) is an importin that transports various cargo proteins into the nucleus through interactions with nucleoporins, which are proteins of the nuclear pore complex (NPC). One might overlook the significance of this protein but it actually plays a crucial role in the human body by mediating transport of RNA-binding proteins involved in transcription, RNA processing, RNA transport and translation. The structure of Kapβ2 is composed of 20 antiparallel helices called HEAT repeats. These HEAT repeats contribute to Kapβ2’s large superhelical shape. The protein is shown to form two arches: one at the N-terminal and the other at the C-terminal. Through recognition of a nuclear localization signal (NLS) located on its cargo, Kapβ2 binds to its cargo via its C-terminal arch. Release of the cargo is mediated by RanGTP, which once bound, modifies the shape of Kapβ2. This slight conformational change results in the dissociation of the cargo from its binding site. The Hostos-Lincoln Academy SMART (Students Modeling a Research Topic) Team and MSOE have designed and made a physical model by three-dimensional (3-D) printing technology, revealing the cargo and RanGTP binding domains. Supported by grants from the HHMI Pre-College Program and the Camille and Henry Dreyfus Foundation.
| | The PY-NLS of 4OO6 contains a basic rather than hydrophobic N-terminal motif. |
| | <scene name='37/372723/Label_nls_arg_glu_interaction/2'>Basic interactions at the N-terminal </scene> motif of the PY-NLS include: Arg92, Arg94, and Arg96 of the NLS with Glu588 and Glu496 of Kapβ2. |
| | <scene name='37/372723/Labelcterminus_interaction_nls/1'>Interactions of the C-terminal</scene> R-X2-5- P-Y motif of the NLS include: Pro98 and Tyr99 of the NLS with Ala381,Ala382, Ala423, Ile457, and Trp460 of Kapβ2. |
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| | Upon binding Kapβ2, the NLS structure conforms to and makes contact with the internal surface of Kapβ2 C-terminal arch |
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| ==='''Background Information'''===
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| [[Image:KapB2_NPC_Schematic.jpg|thumb|alt= Alt text| Figure 1. KapB2 with NLS and cargo moving through the NPC |400px]]
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| In eukaryotes, proteins must be transported in and out of the nucleus. This nucleocytoplasmic transport of proteins across the nuclear envelope must occur through the gateway of the NPC. The NPC is a large structure consisting of 456 constituent binding proteins called nucleoporins (Nups).1 Movement through the NPC is facilitated transport that relies on interaction with specific Nups. Importins and exportins are proteins that aid this facilitated transport by both binding to a specific cargo to be transported and interacting with specific Nups located in the central channel of the NPC.2
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| Karyopherinβ is a group of proteins that is composed of both importins and exportins. Importins are proteins that carry cargos into the nucleus while exportins serve the opposite function. As of today, twenty different Kapβs have been identified. Each of these Kapβs is capable of recognizing and transporting a specific group of cargos. In order to bind to its cargo a Kapβ has to recognize a Nuclear Localizaton or Export Signal (NLS or NES) is located in the polypeptide chain of the cargo. These signals can vary from 7 amino acids to longer than 100 amino acids in length.
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| An importin, such as Kapβ2, binds to a specific cargo by recognition of an NLS and carries the cargo through the NPC by interacting with intrinsically disordered Nups called FG-Nups. FG-Nups line the passageway of the NPC and contain repeats of phenylalanine and glycine. These unstructured FG-Nups form a low-density cloud within the central channel extending from the cytoplasm to the nucleoplasm. The cloud acts as an effective exclusion filter for those particles that do not contain FG repeat binding sites. This is referred to as the zone of selectivity.
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| ==='''How does Kapβ2 identify its cargo?'''===
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| <StructureSection load='2H4M' size=450 side=right scene='2H4M'/Com_view/1'>
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| '''Structure of Kapβ2'''
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| <scene name='Sandbox_122/Kapb2_blue/1'>KapB2</scene>
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| Kapβ2 is a superhelix comprised of 20 HEAT repeats (the name HEAT derives from Huntington, Elongation factor 3 A subunit of protein phosphatase 2A and Tor1 kinase), each of which consists of two <scene name='Sandbox_122/Kapb2_blue_hr2_red/1'>anti-parallel helices.</scene> The electrostatic potential of the internal surface of Kapβ2 superhelix at the C-terminal arch is negative.
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| HEAT repeats <scene name='Sandbox_122/Kapb2_blue_hr9-13_pink/1'>9-13</scene> and <scene name='Sandbox_122/Kapb2_blue_hr14-18_pink/1'>14-18</scene> form the <scene name='Sandbox_122/Kapb2_blue_hr9-13_hr14-18_pink/1'>binding site</scene> of Kapβ2 cargos while repeats <scene name='Sandbox_122/Kapb2_blue_hr1-8_purple/1'>1-8</scene> constitute the RanGTPase binding site.3 RanGTPase, a small 216-residue protein, is found more frequently in the nucleus and enables cargos to be released from Kapβ2.
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| '''Kapβ2 Cargo Binding and Conformational Change4'''
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| The <scene name='Sandbox_122/Kapb2_nls_blue_magenta/3'>NLS</scene> located on Kapβ2 cargos are named the PY- NLS and they bind to the C-terminal arch of Kapβ2.
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| Recognition of the PY-NLS by Kapβ2 follows certain guidelines:
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| (i) PY-NLS, when not bound to Kapβ2, lacks a secondary structure.
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| (ii) PY-NLS has an overall positive charge allowing for electrostatic compatibility with Kapβ2.
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| (iii) General sequence for the PY-NLS is either a hydrophobic or basic motif at the N-terminus and a R-X2-5-P-Y motif at the C-terminus.
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| The <scene name='Sandbox_122/Kapb2_nls_blue_magenta/4'>PY-NLS of 2H4M</scene> contains a hydrophobic rather than a basic N- terminal motif. Hydrophobic interactions at the N- terminal motif of the PY- NLS include: <scene name='Sandbox_122/Kapb2_nls_magenta_hydrophobics/1'>Phe273, Gly274, Pro275 and Met276 </scene>of the NLS with Trp730 and Ile773 of Kapβ2. Interactions at the C- terminal R-X2-5-P-Y motif of the NLS include:
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| <scene name='Sandbox_122/Kapb2_nls_arg284/2'>Arg284</scene> of the NLS with Glu509 and Asp543 of Kapβ2; <scene name='Sandbox_122/Kapb2_nls_pro_tyr/2'>Pro288 and Tyr289</scene> of the NLS with Ala380, Ala 381, Ala 384, Leu419, Ile457, Trp460 and Arg464 of Kapβ2.
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| Upon binding Kapβ2, the NLS gains structure, conforms to and makes contact with the internal surface of the Kapβ2 C-terminal arch. | |
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| '''How does Kapβ2 pass through the NPC?'''
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| Once the cargo binds to Kapβ2, the complex travels through the NPC by interactions with the FG-Nups.
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