Podcasts about Ck1

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Best podcasts about Ck1

Latest podcast episodes about Ck1

PaperPlayer biorxiv cell biology
Fission yeast CK1 promotes DNA double-strand break repair through both homologous recombination and non-homologous end joining

PaperPlayer biorxiv cell biology

Play Episode Listen Later Apr 28, 2023


Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.27.538600v1?rss=1 Authors: Cullati, S. N., Zhang, E., Shan, Y., Guillen, R. X., Chen, J.-S., Navarrete-Perea, J., Elmore, Z. C., Ren, L., Gygi, S. P., Gould, K. L. Abstract: The CK1 family are conserved serine/threonine kinases with numerous substrates and cellular functions. The fission yeast CK1 orthologues Hhp1 and Hhp2 were first characterized as regulators of DNA repair, but the mechanism(s) by which CK1 activity promotes DNA repair had not been investigated. Here, we found that deleting Hhp1 and Hhp2 or inhibiting CK1 catalytic activities in yeast or in human cells activated the DNA damage checkpoint due to persistent double-strand breaks (DSBs). The primary pathways to repair DSBs, homologous recombination and non-homologous end joining, were both less efficient in cells lacking Hhp1 and Hhp2 activity. In order to understand how Hhp1 and Hhp2 promote DSB repair, we identified new substrates using quantitative phosphoproteomics. We confirmed that Arp8, a component of the INO80 chromatin remodeling complex, is a bona fide substrate of Hhp1 and Hhp2 that is important for DSB repair. Our data suggest that Hhp1 and Hhp2 facilitate DSB repair by phosphorylating multiple substrates, including Arp8. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

PaperPlayer biorxiv neuroscience
Regulation of the circadian clock in C. elegans by clock gene homologs kin-20 and lin-42

PaperPlayer biorxiv neuroscience

Play Episode Listen Later Apr 13, 2023


Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.13.536481v1?rss=1 Authors: Lamberti, M. L., Spangler, R. K., Cerdeira, V., Ares, M., Rivollet, L., Ashley, G. E., Coronado, A. R., Tripathi, S., Spiousas, I., Ward, J. D., Partch, C. L., Benard, C. Y., Goya, M. E., Golombek, D. A. Abstract: Circadian rhythms are endogenous oscillations present in nearly all organisms from prokaryotes to humans, allowing them to adapt to cyclical environments close to 24 hours. Circadian rhythms are regulated by a central clock, which is based on a transcription-translation feedback loop. One important protein in the central loop in metazoan clocks is PERIOD, which is regulated in part by Casein kinase 1{varepsilon}/{delta} (CK1{varepsilon}/{delta}) phosphorylation. In the nematode Caenorhabditis elegans, period and casein kinase 1{varepsilon}/{delta} are conserved as lin-42 and kin-20, respectively. Here we studied the involvement of lin-42 and kin-20 in circadian rhythms of the adult nematode using a bioluminescence-based circadian transcriptional reporter. We show that mutations of lin-42 and kin-20 generate a significantly longer endogenous period, suggesting a role for both genes in the nematode circadian clock, as in other organisms. These phenotypes can be partially rescued by overexpression of either gene under their native promoter. Both proteins are expressed in neurons and seam cells, a population of epidermal stem cells in C. elegans that undergo multiple divisions during development. Depletion of LIN-42 and KIN-20 specifically in neuronal cells after development was sufficient to lengthen the period of oscillating sur-5 expression. Therefore, we conclude that LIN-42 and KIN-20 are critical regulators of the adult nematode circadian clock through neuronal cells. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Team Never Quit
Benjamin Sledge: Award Winning Author of Where Cowards Go to Die and Combat Wounded Veteran

Team Never Quit

Play Episode Listen Later Nov 30, 2022 70:00


In this week's Team Never Quit Podcast, Marcus has 11-year combat veteran and Bronze Star and Purple Heart recipient, Benjamin Sledge in the studio. Benjamin served in Iraq and Afghanistan, earning 2 Army Commendation Medals. He lost his best friend in combat. These days, Benjamin is a viral writer, graphic designer, and author of Where Cowards Go to Die where he reveals a brutal portrait of war and the cost of returning to a country that no longer feels like home. He travels around the country educating businesses, non-profits, and churches about veterans' mental health issues. In this episode you will hear: I got to see the best and worst parts of war. Often times we were the first ones in the door, so we either made friends or got shot. I was 21 when I first got to the battle, thinking “I literally have no idea what I'm doing.” There's a very distinct smell to death. It's like rotting meat dabbed with knockoff CK1 cologne. Many combat veterans don't necessarily come home with PTSD; it's moral injury. It's the physiological damage that occurs when you violate your sense of right and wrong. Seeing death from that close does something to the mind. It became a real struggle point for me. When I first got home, they didn't know how to handle me, because I didn't know how to handle myself. I was drinking myself silly, so I could get the images out of my head. The strangest thing happened to me – I found myself missing war.  My wife left me while I was in Iraq. War really is a spiritual experience. What does it look like to live a courageous life and carry that into career and family relationships – Never give up, never accept defeat, and never leave a fallen comrade behind. Inside every man, there's both a warrior and a poet. Support Benjamin Website: https://benjaminsledge.com Book (Where Cowards Go to Die): https://amz.run/5GAb Facebook: https://www.facebook.com/benjamin.sledge/ Instagram: https://www.instagram.com/benjamincsledge/ Twitter: https://twitter.com/benjamincsledge Support TNQ https://www.instagram.com/marcusluttrell/ https://www.instagram.com/andrewbrockenbush/ https://www.instagram.com/team_neverquit/

Bít Tất
Bít Tất #157: Giáo dục giới tính, thế nào là đủ?

Bít Tất

Play Episode Listen Later Jun 18, 2022 23:11


Netflix thì có Sex Education, Vietcetera thì có Cởi Mở, dường như khắp nơi đều đang nói chuyện về giáo dục giới tính. Nhưng là dân F.A lâu năm và có hiểu biết cơ bản về cơ thể mình lẫn quan hệ tình dục, vì sao ta vẫn cần được giáo dục về giới tính? Đồng hành cùng Mai Nguyễn - Editor tại Vietcetera - là ThS. BS. CK1. Trần Quốc Phong, Nhà sáng lập tổ chức truyền thông Saigon Medicine, sẽ giúp bạn nhận ra bạn chưa hiểu về giáo dục giới tính, hay chính bản thân bạn, nhiều như mình tưởng!

Perfume Room
14. Only Virgos Wear Baccarat Rouge (w/ Astrologer David Odyssey)

Perfume Room

Play Episode Listen Later Sep 7, 2021 60:19


In this ep, I am joined by astrologer, reader of the Tarot, and host of The Luminaries podcast, David Odyssey. We talk about the scents each zodiac sign *should* be wearing, traveling to the fifth dimension, the birth charts of your fave (and least fave) perfume-line-having celebrities, our own Big 3 and corresponding scent preferences, and why you MUST AVOID Washington Square Park at all costs. Follow Perfume Room: IG: @perfumeromopod @emmvern TT: @emma_vern Frags mentioned: Witch Oils from Flower Power, Apotheke Rose Bud Oud Candle, Boy Smells Rose Load, Penhaligon's Cairo, Carolina Herrera 212, Escentric Molecules, Tom Ford Lost Cherry, Adam Levine, Fenty, Kierin NYC Rose Ink, Paco Rabanne, CK1, Joop Homme, CK Summer, Dior Sauvage, Baccarat Rouge 540, Creed Aventus, JHAG Not A Perfume, Byredo Gypsy Water, Bleu de Chanel, Le Labo Santal 33, D&G Light Blue, Mugler Alien, YSL Black Opium, TF Black Orchid

Scratch & Sniff
Episode 14 - CK1

Scratch & Sniff

Play Episode Listen Later Jul 30, 2021 3:29


- Scent number 4: CK1 - In this story, memories of adolescence youth from the mid-90s are triggered by the scent of Calvin Klein's unisex cologne, CK1.Music & sound in this episode:90's Rock Style by monkeyman535Link: https://freesound.org/people/monkeyman535/sounds/466981/License: http://creativecommons.org/licenses/by/3.0/Crowded house-ish remix of Fresound #320124.mp3 by TimbreLink: https://freesound.org/people/Timbre/sounds/320196/License: http://creativecommons.org/licenses/by-nc/3.0/Stick swoosh whoosh by HitrisonLink: https://freesound.org/people/Hitrison/sounds/216675/License: http://creativecommons.org/licenses/by/3.0/Bell ding by JakeBaggerLink: https://freesound.org/people/jakebagger/sounds/499787/License: http://creativecommons.org/licenses/by/3.0/

OncLive® On Air
48: FDA Approval Insights: Umbralisib in Marginal Zone Lymphoma and Follicular Lymphoma

OncLive® On Air

Play Episode Listen Later Apr 12, 2021 9:11


In our exclusive interview, Fowler discussed the data that served as the basis for the accelerated approval of umbralisib in marginal zone lymphoma and follicular lymphoma, explained the implications of the regulatory decision on clinical practice, and detailed potential next steps for the PI3Kδ and CK1ε inhibitor.

Don‘t Tread on Merica!
LIVE!! Cancel Culture Corner

Don‘t Tread on Merica!

Play Episode Listen Later Mar 10, 2021 17:31


On todays show! Update on CK1 post surgery! Cancel culture corner!! Short show wanted to check out the going live capabilities!! 

Don‘t Tread on Merica!
The Lost President

Don‘t Tread on Merica!

Play Episode Listen Later Mar 7, 2021 99:16


On todays show PCGC and CK1 discuss "the lost president"! Also is Kamala running the show now? A new name to add to culture cancel corner. And so much more!!     www.facebook.com/donttreadonmerica www.instagram.com/donttreadonmerica www.twitter.com/DTOM_1775  

iBiology Videos
Louis Ptáček & Ying-Hui Fu Part 2: Understanding our Sleep Behaviors

iBiology Videos

Play Episode Listen Later Dec 20, 2020 24:22


By studying families with sleep/wake disorders, Fu and Ptáček have shown that mutations that cause changes in the phosphorylation or acylation of the PER2 protein are responsible for regulating circadian rhythms. In Part 2 of the talk, Dr. Fu explains that studies of families with sleep disorders have shown that post-translational modifications of the PER2 protein are involved in regulating circadian rhythms. Casein kinase 1 phosphorylates PER2 and mutations in either CK1 or specific serine residues in PER2 result in an advanced sleep phase phenotype. O-GlcNAcylation of PER2 also participates in regulating circadian rhythms because O-GlcNAcylation blocks the sites usually phosphorylated by CK1.

PaperPlayer biorxiv neuroscience
The cell-autonomous clock of VIP receptor VPAC2 cells drives circadian behaviour

PaperPlayer biorxiv neuroscience

Play Episode Listen Later Aug 3, 2020


Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.08.02.232462v1?rss=1 Authors: Hamnett, R., Chesham, J. E., Maywood, E. S., Hastings, M. H. Abstract: Circadian (~daily) rhythms pervade mammalian behaviour. They are generated by cell-autonomous, transcriptional/translational feedback loops (TTFL), active in all tissues. This distributed clock network is co-ordinated by the principal circadian pacemaker, the hypothalamic suprachiasmatic nucleus (SCN). Its robust and accurate time-keeping arises from circuit-level interactions that bind its individual cellular clocks into a coherent time-keeper. Cells that express the neuropeptide vasoactive intestinal peptide (VIP) mediate retinal entrainment of the SCN, and in the absence of VIP, or its cognate receptor VPAC2, circadian behaviour is compromised because SCN cells cannot synchronise. The contributions to SCN pacemaking and circadian behaviour of other cell types, not least the VPAC2-expressing target cells of VIP, are, however, not understood. We therefore employed intersectional genetics to manipulate the cell-autonomous TTFL of VPAC2-expressing cells, creating temporally chimaeric mice. We could then determine whether and how VPAC2-expressing cells (a minority ~35% of SCN cells) contribute to SCN time-keeping. Lengthening of the intrinsic TTFL period of VPAC2 cells by deletion of the CK1{varepsilon}Tau allele concomitantly lengthened the period of circadian behavioural rhythms. It also increased the variability of the circadian period of bioluminescent TTFL rhythms in SCN slices recorded ex vivo. Abrogation of circadian competence in VPAC2 cells by deletion of Bmal1 severely disrupted circadian behavioural rhythms and compromised TTFL time-keeping in the corresponding SCN slices. Thus, VPAC2-expressing cells are a distinct, functionally powerful subset of the SCN circuit, contributing to computation of ensemble period and maintenance of circadian robustness. These findings extend our understanding of SCN circuit topology. Copy rights belong to original authors. Visit the link for more info

PaperPlayer biorxiv neuroscience
CHMP2B regulates TDP-43 phosphorylation and proteotoxicity via modulating CK1 turnover independent of the autophagy-lysosomal pathway

PaperPlayer biorxiv neuroscience

Play Episode Listen Later Jun 5, 2020


Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.06.04.133546v1?rss=1 Authors: Sun, X., Deng, X., Hu, R., Duan, Y., Zhang, K., Cui, J., Ni, J., Wang, Q., Chen, Y., Li, A., Fang, Y. Abstract: Protein inclusions containing phosphorylated TDP-43 are a shared pathology in several neurodegenerative diseases including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). However, most ALS/FTD patients do not have a mutation in TDP-43 or the enzymes directly regulating its phosphorylation. It is intriguing how TDP-43 becomes hyperphosphorylated in each disease condition. In a genetic screen for novel TDP-43 modifiers, we found that knockdown (KD) of CHMP2B, a key component of the endosomal sorting complex required for transport (ESCRT) machinery, suppressed TDP-43-mediated neurodegeneration in Drosophila. Further investigation using mammalian cells indicated that CHMP2B KD decreased whereas its overexpression (OE) increased TDP-43 phosphorylation levels. Moreover, a known FTD-causing mutation CHMP2Bintron5 promoted hyperphosphorylation, insolubility and cytoplasmic accumulation of TDP-43. Interestingly, CHMP2B did not manifest these effects by its well-known function in the autophagy-lysosomal pathway. Instead, the kinase CK1 tightly regulated TDP-43 phosphorylation level in cells, and CHMP2B OE or CHMP2BIntron5 significantly decreased ubiquitination levels and the turnover of CK1 via the ubiquitin-proteasome (UPS) pathway. Finally, we showed that CHMP2B protein levels increased in the cerebral cortices of aged mice, which might underlie the age-associated TDP-43 pathology and disease onset. Together, our findings reveal a molecular link between the two ALS/FTD-pathogenic proteins CHMP2B and TDP-43, and provide an autophagy-independent mechanism for CHMP2B in pathogenesis. Copy rights belong to original authors. Visit the link for more info

Bagateller
#22 Gnu i bakrus, tannlegen og 90 talls parfymer

Bagateller

Play Episode Listen Later Feb 25, 2020 43:39


Henrik har hatt en drømmedag; han har vært hos tannlegen og betalt 400 spenn mer enn han pleier. Jannecke skjønner ikke hvorfor noen gidder å bli tannlege, men vurderer å bli sykepleier og ha noe å falle tilbake på. Men hun er ikke et sånt godt menneske. Jannecke blir gnu dagen derpå, og angrer alltid på at hun har drukket, mens Henrik har noen av sine beste dager i bakrus. I nostalgihjørnet denne gangen handler det om parfymer fra 90 tallet. Date, Aqua di Gio, Lagerfeld og CK1. Noen lukter hodepine og er døde nå, mens andre vekker søte minner fra ungdomsskolen. See acast.com/privacy for privacy and opt-out information.

First laugh podcast
#62: Ouija board comedy w/ Nicholas Cuvar

First laugh podcast

Play Episode Listen Later Feb 24, 2020 80:49


On this episode I have a candlelit chat with St Louis comic Nicholas Cuvar. We discuss summoning the spirits, independent comedy, and much much more!   Like share prescribe! 

90's Made Me: Music + Nostalgia Podcast
"Licky Boom Boom Down", Jim Carrey's Parody "Imposter", and Caboodles

90's Made Me: Music + Nostalgia Podcast

Play Episode Listen Later Apr 17, 2019 10:38


Meredith digs in on Snow's "Informer" from 1992, Jim Carrey's parody "Imposter", and a 2019 sample with Daddy Yankee. She talks caboodles and scents of the early '90s.

High Performance Mindset | Learn from World-Class Leaders, Consultants, Athletes & Coaches about Mindset
198: How to Build and Sustain a Championship Culture with Jeff Janssen, Janssen Sport Leadership Center, Founder and Director

High Performance Mindset | Learn from World-Class Leaders, Consultants, Athletes & Coaches about Mindset

Play Episode Listen Later Jul 28, 2018 54:26


Widely considered the world's top expert on Sports Leadership, Jeff Janssen is the founder and president of the Janssen Sports Leadership Center. Jeff's pioneering work in launching and conducting college athletics' first Leadership Academies with student-athletes and coaches began over a decade ago. His groundbreaking work continues to be the gold standard today. Jeff and his team's work has led to the creation of cutting edge Leadership Academies at Michigan, Illinois, Arkansas, LSU, Pitt, NC State, Colorado, Yale, Georgetown, South Carolina, Stanford, among others. He has authored numerous books including The Team Captain's Leadership Manual, The Team Captain's Culture Manual, Championship Team Building, Jeff Janssen's Peak Performance Playbook, How to Develop Relentless Competitors, The Commitment Continuum™ System, and The Seven Secrets of Successful Coaches. In this interview, Jeff and Cindra talk about: 6 key components in a championship culture The characteristics of a high performance culture 7 secrets of successful coaches How and why he developed the leadership academies How to build and sustain a championship culture[CK1]  10 step blueprint You can find a full description of the Podcast at cindrakamphoff.com/jeffjanssen

80 of the 80s Music Podcast
#64: Olivia Newton-John - Twist of Fate (& Workout Songs)

80 of the 80s Music Podcast

Play Episode Listen Later Nov 25, 2015 49:31


They're two of a kind - featuring a double-dose of upbeat, power-packed, energy-fueled commentary and a sprinkle of torturous laughter. Even if AK47 is not A.2.O.N.J., even if CK1 can't pronounce "Farrar," and he's drinking Jimmy Buffett beer, and the Gobotron 2000 ain't working - our montage of montages will release the twist of fate that will save humanity! Links: Our 80s Workout Song YouTube Playlist CBC's List of 10 Best 80s Training Montage Songs Intellivisionaries (intellivisionaries.com) Throwback Reviews (throwbackreviews.com)

80 of the 80s Music Podcast
#66: The Go-Go's - Head Over Heels (& Kraftwerk Concert Review)

80 of the 80s Music Podcast

Play Episode Listen Later Oct 10, 2015 38:31


Which Go-Go does CK1 go for? AK-47 experiences 3 dimensions of Kraftwerk. Will she keep her cool? Which version of Our Lips Are Sealed is more fun? More homicidal?

80 of the 80s Music Podcast
#68: Thompson Twins - Doctor! Doctor!

80 of the 80s Music Podcast

Play Episode Listen Later Aug 19, 2015 25:44


In an episode full of contradictions, it’s Mullet vs. Mohawk vs. Rattail. AK-47 attempts to keep things PG. CK1 admits to a burning sensation and a Pac-Man sleeping bag. Other mentioned artists: Wham, Babble, Weird Al, Kate Bush, Hall & Oats, Wire, Perry Farrell, Robey, Laura Branigan, After The Fire, Falco

80 of the 80s Music Podcast
#69: Murray Head - One Night in Bangkok

80 of the 80s Music Podcast

Play Episode Listen Later Jul 22, 2015 23:29


Where does Mr. Head get his kicks?  Will the glowing heads elect the Arbiter as Dictator of the World?  Can you hear the winking in his voice?  Reagan gives Gorbachev a wedgie.  What beer does CK1 prefer?Other mentioned artists: Yaz/Yazoo, Anders Glenmark, Björn Skifs, Abba, Elaine Paige, Barbara Dickson, Pet Shop Boys, David Byrne, M, Zamfir, David Hasselhoff.

80 of the 80s Music Podcast
#72: The Stone Roses - She Bangs the Drums

80 of the 80s Music Podcast

Play Episode Listen Later May 19, 2015 20:11


Tired of CK1's incessant rambling, AK-47 takes control of the mic. We learn of the Brit-Pop Triangle of Hate, the importance of pelvic floor exercises, and the tragic consequences of sneak peaks. Featuring a special guest video review! Other mentioned artists: Vixen, Suede, Oasis, Blur, Gin Blossoms, Elvis, Menudo, Kraftwerk (In 3-D)

Probably Science
Episode 109 - Tara Flynn

Probably Science

Play Episode Listen Later Apr 1, 2014 90:56


Irish comedian, actress and voice artist Tara Flynn (twitter.com/TaraFlynn) joins Matt, Jesse and Andy this week to discuss: Fleet Week! Sex workers in the Pacific Northwest! Sharing a name with a porn star! Jesse spraying himself with urine before a callback! The Shewee! Saving the government $400 million by changing fonts! Ink cartels! Debunking St. Patrick! Carving off a sliver of Irish Spring soap! A MacGyver microscope that only costs fifty cents! The possibly bullshit phenomenon of autonomous sensory meridian response! A 3D-printed skull! Tara's lack of a sense of smell! The influence of CK1 (not the perfume) on our circadian rhythms! Graphene contact lenses that let you see in the dark! Andy's almost revolutionary inventions!

Above the Noise
#21 ATN Interview with Hit Writer, Producer, Publisher & Radio Promoter, Mike James

Above the Noise

Play Episode Listen Later Aug 21, 2012 36:59


After being away a few weeks getting married and spending time with family and friends I am back with a new episode of "Above The Noise." This week I have Mike James on the show and he has a ton of information to share! From writing Billboard hits to radio promoting, music publishing and music placements, Mike has a great wealth of music industry knowledge! He is the owner of Milk Music Co., a publishing company administered by Netwerk One Music, and co-founder and president of HipJoint, a production company and label with distribution via Universal Music. Mike has had success with artists of all styles. Kelly Rowland "Unity" - released worldwide on her Diva Deluxe album - Top 10 radio hit in Canada, Delerium "Dust In Gravity" - Hipjoint were the top line writers on this track that achieved US Billboard #1 status on the Dance/Club Play chart in 2010, Calvin Klein CK1 - Hipjoint's song "We Are One" was used as the positioning track for Calvin Klein's worldwide relaunch of CK1 in 2009 Mika Nakashima "Candy Girl" - The Japanese superstar recently took this Hipjoint track to #4 on Japan's Oricon Weekly Singles Chart, Shiloh "Operator (A Girl Like Me)" - Hipjoint discovered this Canadian artist at 13 and wrote and produced her platinum debut hit for her (Universal Music Canada/Universal Republic), Nikon Cool Pix - Nikon used Hipjoint's track "Bounce With Me" by Kreesha Turner as it's positioning song for a series of Nikon Hip Joint Productions Mike JamesCool Pix ads featuring Ashton Kutcher, NFL Network - The NFL has chosen the Hipjoint written and produced "Crank It Up" by The Young Goulets to promote 2010 NFL Thursday Night Football. With success at writing hits, promoting singles on the radio, placing music in TV and film, as well as developing his businesses I have asked Mike what he does to be heard above the noise in all that he does! Aaron Bethune. In our conversation we discuss: What makes a hit Writing for radio Music discovery Finding new talent Music placements What are the parts that make up a successful artist? What is success? How you get heard "Above The Noise."

Autoline Daily - Video
Episode 508 - Exchange Rate Crisis, Geely Receives Zero Stars, 007's Aston Up for Auction

Autoline Daily - Video

Play Episode Listen Later Oct 27, 2010 7:35


Nissan says that every one yen increase against the dollar shaves nearly $190 million off its profits. The CK1 from Chinese automaker Geely received zero stars in crash tests. A 1964 Aston Martin DB5 driven by Sean Connery in the James Bond movies "Goldfinger" and "Thunderball" is up for auction. All that and more, plus John responds to your comments in the "You Said It!" segment.

Fakultät für Chemie und Pharmazie - Digitale Hochschulschriften der LMU - Teil 01/06

Die Inhibition der b-Sekretase stellt derzeit einen vielversprechenden Ansatz zur Therapie der Alzheimer Krankheit dar. Der Hauptanteil der b-Sekretase Aktivität ist auf BACE-1, eine neuartige membrangebundene Typ I Aspartylprotease, zurückzuführen. Um gezielt spezifische und wirksame Inhibitoren entwickeln zu können, ist es notwendig, die Eigenschaften und katalytischen Spezifitäten der Protease genauer zu verstehen. Da neben BACE-1 eine weitere hochgradig homologe Aspartylprotease, BACE-2, bekannt ist, ist es für die Suche nach Inhibitoren ferner von Bedeutung, charakteristische Unterschiede zwischen den beiden Enzymen zu kennen, um mögliche Kreuzreaktionen der Inhibitoren minimieren zu können. Ziel dieser Arbeit war es deshalb, die beiden Enzyme bezüglich ihrer posttranslationalen Modifikationen und insbesondere ihrer katalytischen Spezifitäten vergleichend zu analysieren. Als Modell für die durchgeführten Experimente dienten HEK293 Zellen, mit exogener Expression der beiden Proteasen, sowie dem Substrat bAPP. Beide Proteine werden in ähnlicher Weise durch die kovalente Bindung komplexer Kohlehydrateinheiten modifiziert. Matures BACE-1 besitzt im Vergleich zu BACE-2 eine eine längere Halbwertszeit. In vitro werden beide Enzyme durch CK1 an homologen Serinen in der zytoplasmatischen Domäne phosphoryliert. Während für BACE-2 bisher nicht schlüssig gezeigt werden konnte, dass auch in vivo eine Phosphorylierung erfolgt, wurde für BACE-1 S498 auch als Phosphorylierungsstelle in vivo bestätigt. Mittels Biotinylierung konnte demonstriert werden, dass beide BACE-Proteasen effizient an die Zelloberfläche transportiert werden. Im Gegensatz zu BACE-1, welches rasch in endosomale Kompartimente reinternalisiert wird und phosphorylierungsabhängig zurück zum TGN transportiert wird, wird BACE-2 entweder durch Spaltung der Ektodomäne in den extrazellulären Raum sezerniert, oder aber unmittelbar nach der Reinternalsierung ins Zellinnere in lysosomalen Kompartimenten abgebaut. Dieser Unterschied begründet vermutlich die unterschiedlichen Halbwertszeiten der beiden Proteine und erhöht gleichzeitig die Gesamtverweildauer von BACE-1 in endosomalen Kompartimenten, die aufgrund ihres pH-Wertes günstige Bedingungen für die proteolytische Aktivität des Enzyms schaffen. Hinsichtlich der katalytischen Spezifität bezüglich des membrangebundenen bAPP unterscheiden sich BACE-1 und BACE-2 grundlegend. Während BACE-1 die erwarteten b-Sekretase Spaltungen an Asp1 und Glu11 der Ab-Domäne katalysiert, spaltet BACE-2 vorzugsweise zwischen Phe19 und Phe20 der Ab-Domäne, wodurch Spaltprodukte entstehen, die denen der a-Sekretase Spaltung ähneln. Durch Koexpression der beiden Enzyme konnte gezeigt werden, dass BACE-2 die BACE-1 abhängige Prozessierung des Substrates direkt oder indirekt beeinflussen kann. Die Behandlung der entsprechenden Zelllinien mit BFA oder Monensin belegt, dass BACE-1 bereits in den frühen Kompartimenten des sekretorischen Prozessierungsweges proteolytisch aktiv sein kann, während BACE-2 auch nach exogener Expression keine Aktivität in diesen Kompartimenten zeigt. Mit Hilfe massenspektrometrischer Analysen wurde bewiesen, dass BACE-1 und BACE-2 entgegen bisheriger Annahmen nicht ausschließlich die proteolytische Spaltung membrangebundener bAPP Substrate katalysieren, sondern zudem Ab-Peptide, nach ihrer Freisetzung durch g-Sekretase, C-terminal verkürzen können. In vitro Versuche zeigen, dass BACE-1 selbst in der Lage ist, Ab 1-40 an Position 34 zu spalten und dieser Schnitt nicht wie bislang angenommen durch g-Sekretase katalysiert wird. Dieser Vorgang führt in vivo zu einer Reduktion der amyloidogenen Ab 1-40/42 Peptide. Da sich der Nachweis des Ab 1-34 Peptides mittels konservativer Proteinanalytik schwierig gestaltet, erklärt sich, warum in Zelllinien mit exogener BACE-1 Expression keine merkliche Steigerung der Ab-Sezernierung bzw. teilweise sogar eine Reduktion detektierbar war. Letztendlich bietet die Beobachtung, dass auch Peptide als Substrate für die BACE fungieren können, interessante Ansatzpunkte für die Suche nach neuen physiologischen Substraten und Inhibitoren. Die Analyse des subzellulären Transportes und die Charakterisierung, sowohl pro- als auch antiamyloidogener Enzymaktivitäten der beiden Proteasen BACE-1 und BACE-2 liefert neue Grundlagen für die Entwicklung therapeutischer Inhibitoren und für die Suche neuer Substrate.

position arbeit ab dom ziel entwicklung suche raum bedeutung weise lage expression zwei unterschiede vergleich ansatz bfa bedingungen aktivit therapie eigenschaften grundlagen mit hilfe schnitt experimente versuche analysen bindung beobachtung im gegensatz peptides spaltung steigerung transportes enzyme reduktion annahmen proteine letztendlich nachweis die analyse substrate ansatzpunkte die behandlung hinsichtlich charakterisierung freisetzung halbwertszeit protease spezifit modifikationen enzymen bace tgn inhibitoren dieser vorgang ddc:500 zelllinien als modell phosphorylierung enzyms alzheimer krankheit zelloberfl prozessierung ck1 enzymaktivit dieser unterschied substraten ddc:540 proteasen ph wertes kreuzreaktionen sekretase kompartimente hek293 zellen kompartimenten monensin die inhibition bapp beide proteine
The Baller Lifestyle Podcast
EP. 242: No Nut November

The Baller Lifestyle Podcast

Play Episode Listen Later Jan 1, 1970 100:35


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