Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor

Fytokromit ovat punaista ja kaukopunaista valoa aistivia valosensoriproteiineja, jotka säätelevät valosta riippuvaisia prosesseja niin kasveissa, sienissä kuin bakteereissakin. Niillä on kaksi konformaatiota, Pfr- ja Pr-tila, joiden välillä ne vaihtavat reagoidessaan punaiseen ja kaukopunaiseen valo...

Full description

Bibliographic Details
Main Author: Jokelainen, Kaisla
Other Authors: Matemaattis-luonnontieteellinen tiedekunta, Faculty of Sciences, Bio- ja ympäristötieteiden laitos, Department of Biological and Environmental Science, Jyväskylän yliopisto, University of Jyväskylä
Format: Master's thesis
Language:eng
Published: 2025
Subjects:
Online Access: https://jyx.jyu.fi/handle/123456789/103884
_version_ 1836034467467100160
author Jokelainen, Kaisla
author2 Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä
author_facet Jokelainen, Kaisla Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä Jokelainen, Kaisla Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä
author_sort Jokelainen, Kaisla
datasource_str_mv jyx
description Fytokromit ovat punaista ja kaukopunaista valoa aistivia valosensoriproteiineja, jotka säätelevät valosta riippuvaisia prosesseja niin kasveissa, sienissä kuin bakteereissakin. Niillä on kaksi konformaatiota, Pfr- ja Pr-tila, joiden välillä ne vaihtavat reagoidessaan punaiseen ja kaukopunaiseen valoon. Bakteerifytokromit jaetaan kanonisiin ja bathy-fytokromeihin sen perusteella, mikä konformaatiotila on vakaa pimeässä. Kanonisilla fytokromeilla vakaa tila on Pr, kun taas bathy-fytokromeilla se on Pfr. Tässä tutkielmassa karakterisoitiin Sphingomonas glacialis -bakteerin fytokromi SgBphP1. Fytokromi puhdistettiin onnistuneesti jatkotutkimuksia varten. SgBphP1 määritettiin dimeeriseksi sekä punaisella että kaukopunaisella valolla valotettaessa. Sen aggregoitumista saatiin vähennettyä käyttämällä 2 mM DTT:tä. UV-vis-spektroskopia osoitti fotokonversion molempiin suuntiin, mutta punainen valo tuotti Pfr- ja Pr-tilan sekoituksen. SgBphP1 asettui +7 °C:n ja +25 °C:n välillä Pr- ja Pfr-tilan sekoitukseen, ja pimeäpalautuminen nopeutui sitä enemmän, mitä lämpimämmässä fytokromia pidettiin. Koska kumpikaan tila ei ollut yksinomainen pimeätila, SgBphP1 edustaa välimuotoa kanonisten ja bathyfytokromien välillä ja sijoittuu niin sanottuun bathy-like-alaluokkaan. Tämä selvitys SgBphP1:n fotokemiasta luo pohjan sen signaloinnin tutkimiselle sekä sen selvittämiselle, mitkä rakenteelliset ominaisuudet aiheuttavat sen välityyppisen tasapainotilan. Phytochromes convert red/far-red light into biochemical signals that regulate physiological processes in plants, bacteria, and fungi. They switch between two conformations, a red light absorbing Pr state and a far-red light absorbing Pfr state, as a response to light. Depending on which state is the most stable in darkness, bacterial phytochromes are classified as prototypical (Pr ground state) or bathy (Pfr ground state). Here, I characterised SgBphP1, a bacteriophytochrome from Sphingomonas glacialis, to test how temperature affects its thermal reversion. SgBphP1 was determined to be dimeric under both red and far-red light, and aggregation was reduced with 2 mM DTT. UV-vis spectroscopy showed bidirectional photoconversion, but red light created only a partial accumulation of Pfr state. Thermal reversion assay in temperatures from +7 °C to +25 °C revealed a faster reversion speed at higher temperatures and a slightly higher Pfr:Pr ratio at lower temperatures. The end-point spectrum remained a mixed Pfr:Pr state, implying that neither state was an absolute ground state. The spectral characteristics place SgBphP1 between prototypical and bathy phytochromes to an intermediate bathy-like group, which is relatively unexplored. These initial photochemical data provide a foundation for testing whether SgBphP1 output module functions as a histidine kinase or phosphatase and mapping the molecular bases of the bathy-like ground state equilibrium.
first_indexed 2025-06-23T20:00:37Z
format Pro gradu
fullrecord [{"key": "dc.contributor.advisor", "value": "Ihalainen, Janne", "language": null, "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Rumfeldt, Jessica", "language": null, "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Jokelainen, Kaisla", "language": null, "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2025-06-23T12:35:14Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2025-06-23T12:35:14Z", "language": null, "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2025", "language": null, "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/103884", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Fytokromit ovat punaista ja kaukopunaista valoa aistivia valosensoriproteiineja, jotka s\u00e4\u00e4telev\u00e4t valosta riippuvaisia prosesseja niin kasveissa, sieniss\u00e4 kuin bakteereissakin. Niill\u00e4 on kaksi konformaatiota, Pfr- ja Pr-tila, joiden v\u00e4lill\u00e4 ne vaihtavat reagoidessaan punaiseen ja kaukopunaiseen valoon. Bakteerifytokromit jaetaan kanonisiin ja bathy-fytokromeihin sen perusteella, mik\u00e4 konformaatiotila on vakaa pime\u00e4ss\u00e4. Kanonisilla fytokromeilla vakaa tila on Pr, kun taas bathy-fytokromeilla se on Pfr. T\u00e4ss\u00e4 tutkielmassa karakterisoitiin Sphingomonas glacialis -bakteerin fytokromi SgBphP1. Fytokromi puhdistettiin onnistuneesti jatkotutkimuksia varten. SgBphP1 m\u00e4\u00e4ritettiin dimeeriseksi sek\u00e4 punaisella ett\u00e4 kaukopunaisella valolla valotettaessa. Sen aggregoitumista saatiin v\u00e4hennetty\u00e4 k\u00e4ytt\u00e4m\u00e4ll\u00e4 2 mM DTT:t\u00e4. UV-vis-spektroskopia osoitti fotokonversion molempiin suuntiin, mutta punainen valo tuotti Pfr- ja Pr-tilan sekoituksen. SgBphP1 asettui +7 \u00b0C:n ja +25 \u00b0C:n v\u00e4lill\u00e4 Pr- ja Pfr-tilan sekoitukseen, ja pime\u00e4palautuminen nopeutui sit\u00e4 enemm\u00e4n, mit\u00e4 l\u00e4mpim\u00e4mm\u00e4ss\u00e4 fytokromia pidettiin. Koska kumpikaan tila ei ollut yksinomainen pime\u00e4tila, SgBphP1 edustaa v\u00e4limuotoa kanonisten ja bathyfytokromien v\u00e4lill\u00e4 ja sijoittuu niin sanottuun bathy-like-alaluokkaan. T\u00e4m\u00e4 selvitys SgBphP1:n fotokemiasta luo pohjan sen signaloinnin tutkimiselle sek\u00e4 sen selvitt\u00e4miselle, mitk\u00e4 rakenteelliset ominaisuudet aiheuttavat sen v\u00e4lityyppisen tasapainotilan.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Phytochromes convert red/far-red light into biochemical signals that regulate physiological processes in plants, bacteria, and fungi. They switch between two conformations, a red light absorbing Pr state and a far-red light absorbing Pfr state, as a response to light. Depending on which state is the most stable in darkness, bacterial phytochromes are classified as prototypical (Pr ground state) or bathy (Pfr ground state). Here, I characterised SgBphP1, a bacteriophytochrome from Sphingomonas glacialis, to test how temperature affects its thermal reversion. SgBphP1 was determined to be dimeric under both red and far-red light, and aggregation was reduced with 2 mM DTT. UV-vis spectroscopy showed bidirectional photoconversion, but red light created only a partial accumulation of Pfr state. Thermal reversion assay in temperatures from +7 \u00b0C to +25 \u00b0C revealed a faster reversion speed at higher temperatures and a slightly higher Pfr:Pr ratio at lower temperatures. The end-point spectrum remained a mixed Pfr:Pr state, implying that neither state was an absolute ground state. The spectral characteristics place SgBphP1 between prototypical and bathy phytochromes to an intermediate bathy-like group, which is relatively unexplored. These initial photochemical data provide a foundation for testing whether SgBphP1 output module functions as a histidine kinase or phosphatase and mapping the molecular bases of the bathy-like ground state equilibrium.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by Miia Hakanen (mihakane@jyu.fi) on 2025-06-23T12:35:14Z\nNo. of bitstreams: 0", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2025-06-23T12:35:14Z (GMT). No. of bitstreams: 0\n Previous issue date: 2025", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "49", "language": null, "element": "format", "qualifier": "extent", "schema": "dc"}, {"key": "dc.language.iso", "value": "eng", "language": null, "element": "language", "qualifier": "iso", "schema": "dc"}, {"key": "dc.rights", "value": "In Copyright", "language": null, "element": "rights", "qualifier": null, "schema": "dc"}, {"key": "dc.subject.other", "value": "Bacteriophytochrome", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "bathy-like photoreceptor", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "dimerisation", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "photoconversion", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "photo state equilibrium", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "SgBphP1", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "thermal reversion", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.title", "value": "Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor", "language": null, "element": "title", "qualifier": null, "schema": "dc"}, {"key": "dc.type", "value": "master thesis", "language": null, "element": "type", "qualifier": null, "schema": "dc"}, {"key": "dc.identifier.urn", "value": "URN:NBN:fi:jyu-202506235648", "language": null, "element": "identifier", "qualifier": "urn", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Matemaattis-luonnontieteellinen tiedekunta", "language": "fi", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Faculty of Sciences", "language": "en", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.department", "value": "Bio- ja ymp\u00e4rist\u00f6tieteiden laitos", "language": "fi", "element": "contributor", "qualifier": "department", "schema": "dc"}, {"key": "dc.contributor.department", "value": "Department of Biological and Environmental Science", "language": "en", "element": "contributor", "qualifier": "department", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "Jyv\u00e4skyl\u00e4n yliopisto", "language": null, "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "University of Jyv\u00e4skyl\u00e4", "language": null, "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Solu- ja molekyylibiologia", "language": "fi", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Cell and molecular biology", "language": "en", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "dc.type.coar", "value": "http://purl.org/coar/resource_type/c_bdcc", "language": null, "element": "type", "qualifier": "coar", "schema": "dc"}, {"key": "dc.rights.copyright", "value": "\u00a9 The Author(s)", "language": "fi", "element": "rights", "qualifier": "copyright", "schema": "dc"}, {"key": "dc.rights.accesslevel", "value": "restrictedAccess", "language": null, "element": "rights", "qualifier": "accesslevel", "schema": "dc"}, {"key": "dc.type.publication", "value": "masterThesis", "language": null, "element": "type", "qualifier": "publication", "schema": "dc"}, {"key": "dc.subject.yso", "value": "spektroskopia", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "fytokromit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "l\u00e4mp\u00f6tila", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.rights.url", "value": "https://rightsstatements.org/page/InC/1.0/", "language": null, "element": "rights", "qualifier": "url", "schema": "dc"}, {"key": "dc.rights.accessrights", "value": "Tekij\u00e4 ei ole antanut lupaa avoimeen julkaisuun, joten aineisto on luettavissa vain Jyv\u00e4skyl\u00e4n yliopiston kirjaston arkistoty\u00f6semalta. Ks. https://kirjasto.jyu.fi/fi/tyoskentelytilat/laitteet-ja-tilat#autotoc-item-autotoc-2", "language": "fi", "element": "rights", "qualifier": "accessrights", "schema": "dc"}, {"key": "dc.rights.accessrights", "value": "The author has not given permission to make the work publicly available electronically. Therefore the material can be read only at the archival workstation at Jyv\u00e4skyl\u00e4 University Library (https://kirjasto.jyu.fi/fi/tyoskentelytilat/laitteet-ja-tilat#autotoc-item-autotoc-2.", "language": "en", "element": "rights", "qualifier": "accessrights", "schema": "dc"}]
id jyx.123456789_103884
language eng
last_indexed 2025-06-23T20:01:32Z
main_date 2025-01-01T00:00:00Z
main_date_str 2025
publishDate 2025
record_format qdc
source_str_mv jyx
spellingShingle Jokelainen, Kaisla Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor Bacteriophytochrome bathy-like photoreceptor dimerisation photoconversion photo state equilibrium SgBphP1 thermal reversion Solu- ja molekyylibiologia Cell and molecular biology spektroskopia fytokromit lämpötila
title Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor
title_full Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor
title_fullStr Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor
title_full_unstemmed Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor
title_short Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor
title_sort characterisation of sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor
title_txtP Characterisation of Sphingomonas glacialis phytochrome reveals an intermediate type between bathy and prototypical photosensor
topic Bacteriophytochrome bathy-like photoreceptor dimerisation photoconversion photo state equilibrium SgBphP1 thermal reversion Solu- ja molekyylibiologia Cell and molecular biology spektroskopia fytokromit lämpötila
topic_facet Bacteriophytochrome Cell and molecular biology SgBphP1 Solu- ja molekyylibiologia bathy-like photoreceptor dimerisation fytokromit lämpötila photo state equilibrium photoconversion spektroskopia thermal reversion
url https://jyx.jyu.fi/handle/123456789/103884 http://www.urn.fi/URN:NBN:fi:jyu-202506235648
work_keys_str_mv AT jokelainenkaisla characterisationofsphingomonasglacialisphytochromerevealsanintermediatetypebetw