Protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in Antarctic fish
Temperature dependent growth is an important indicator to understand the thermal tolerance of organisms and to project their vulnerability to future climate change. Direct measurements of temperature dependent weight gains, however, are experimentally challenging and time consuming in long-lived spe...
Main Authors: | , , , , , , |
---|---|
Format: | Dataset |
Language: | English |
Published: |
PANGAEA
2022
|
Subjects: | |
Online Access: | https://doi.pangaea.de/10.1594/PANGAEA.940626 https://doi.org/10.1594/PANGAEA.940626 |
id |
ftpangaea:oai:pangaea.de:doi:10.1594/PANGAEA.940626 |
---|---|
record_format |
openpolar |
spelling |
ftpangaea:oai:pangaea.de:doi:10.1594/PANGAEA.940626 2024-09-15T17:46:38+00:00 Protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in Antarctic fish Krebs, Nina Tebben, Jan Pörtner, Hans-Otto Lannig, Gisela Lucassen, Magnus Mark, Felix Christopher Bock, Christian LATITUDE: 53.533000 * LONGITUDE: 8.580100 2022 text/tab-separated-values, 1993 data points https://doi.pangaea.de/10.1594/PANGAEA.940626 https://doi.org/10.1594/PANGAEA.940626 en eng PANGAEA Krebs, Nina; Bock, Christian; Tebben, Jan; Mark, Felix Christopher; Lucassen, Magnus; Lannig, Gisela; Pörtner, Hans-Otto (2023): Evolutionary Adaptation of Protein Turnover in White Muscle of Stenothermal Antarctic Fish: Elevated Cold Compensation at Reduced Thermal Responsiveness [dataset]. PANGAEA, https://doi.org/10.1594/PANGAEA.963276 https://doi.pangaea.de/10.1594/PANGAEA.940626 https://doi.org/10.1594/PANGAEA.940626 UNKNOWN: Licensing unknown: Please contact principal investigator/authors to gain access and request licensing terms Access constraints: access rights needed info:eu-repo/semantics/restrictedAccess 13C-labeling Antarctica AWI_lab Bremerhaven Germany Calculated cold adaptation Energy budget EXP Experiment Hepato-somatic index Liquid chromatography-high resolution mass spectrometry (LC-HRMS) with Orbitrap Pachycara brachycephalum liver mass standard length Phenylalanine labelled in cytosol area in protein unlabelled physiology Proteins dry mass synthesis rate per day Sample wet mass Sample code/label Sample code/label 2 Scale Subsample ID dataset 2022 ftpangaea https://doi.org/10.1594/PANGAEA.94062610.1594/PANGAEA.963276 2024-07-24T02:31:34Z Temperature dependent growth is an important indicator to understand the thermal tolerance of organisms and to project their vulnerability to future climate change. Direct measurements of temperature dependent weight gains, however, are experimentally challenging and time consuming in long-lived species. Here, we reassess methodology to quantify the in vivo protein synthesis rate from amino acids, as a key component of growth. We developed an analytical method that is both robust against analytical errors and does not require hazardous radioactive materials. We utilized the incorporation of isotopically 13C15N-labeled-phenylalanine into fish muscle followed by quantification by liquid chromatography mass spectrometry to calculate accurate net protein synthesis rates in muscle tissue of Antarctic fish, Pachycara brachycephalum, in vivo. Specifically, we injected 150 mM of 13C9H915N1 phenylalanine intraperitoneally and sampled muscle tissue in 1,5h steps between 0 and 6 hours after injection. We quantified labeled and unlabeled phenylalanine both in muscle protein and in the cytosol. This allowed us to critically re-evaluate three different protein synthesis rate (Ks) calculation methodologies that have been developed over the last decades. The calculated values differ by more than 70-fold (0.048 ± 0.021% day-1 up to 3.56 ± 2.16 day-1) between methods. We argue that the Ks calculation including a proportionate ratio of protein synthesis from (unlabeled) free amino acids yields the most realistic Ks values for cold water fish. Eventually, the standardization of the net protein synthesis rate calculation will lead to dependable quantitative representations of organismal stress in response to climate change. Dataset Antarc* Antarctic Antarctica PANGAEA - Data Publisher for Earth & Environmental Science ENVELOPE(8.580100,8.580100,53.533000,53.533000) |
institution |
Open Polar |
collection |
PANGAEA - Data Publisher for Earth & Environmental Science |
op_collection_id |
ftpangaea |
language |
English |
topic |
13C-labeling Antarctica AWI_lab Bremerhaven Germany Calculated cold adaptation Energy budget EXP Experiment Hepato-somatic index Liquid chromatography-high resolution mass spectrometry (LC-HRMS) with Orbitrap Pachycara brachycephalum liver mass standard length Phenylalanine labelled in cytosol area in protein unlabelled physiology Proteins dry mass synthesis rate per day Sample wet mass Sample code/label Sample code/label 2 Scale Subsample ID |
spellingShingle |
13C-labeling Antarctica AWI_lab Bremerhaven Germany Calculated cold adaptation Energy budget EXP Experiment Hepato-somatic index Liquid chromatography-high resolution mass spectrometry (LC-HRMS) with Orbitrap Pachycara brachycephalum liver mass standard length Phenylalanine labelled in cytosol area in protein unlabelled physiology Proteins dry mass synthesis rate per day Sample wet mass Sample code/label Sample code/label 2 Scale Subsample ID Krebs, Nina Tebben, Jan Pörtner, Hans-Otto Lannig, Gisela Lucassen, Magnus Mark, Felix Christopher Bock, Christian Protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in Antarctic fish |
topic_facet |
13C-labeling Antarctica AWI_lab Bremerhaven Germany Calculated cold adaptation Energy budget EXP Experiment Hepato-somatic index Liquid chromatography-high resolution mass spectrometry (LC-HRMS) with Orbitrap Pachycara brachycephalum liver mass standard length Phenylalanine labelled in cytosol area in protein unlabelled physiology Proteins dry mass synthesis rate per day Sample wet mass Sample code/label Sample code/label 2 Scale Subsample ID |
description |
Temperature dependent growth is an important indicator to understand the thermal tolerance of organisms and to project their vulnerability to future climate change. Direct measurements of temperature dependent weight gains, however, are experimentally challenging and time consuming in long-lived species. Here, we reassess methodology to quantify the in vivo protein synthesis rate from amino acids, as a key component of growth. We developed an analytical method that is both robust against analytical errors and does not require hazardous radioactive materials. We utilized the incorporation of isotopically 13C15N-labeled-phenylalanine into fish muscle followed by quantification by liquid chromatography mass spectrometry to calculate accurate net protein synthesis rates in muscle tissue of Antarctic fish, Pachycara brachycephalum, in vivo. Specifically, we injected 150 mM of 13C9H915N1 phenylalanine intraperitoneally and sampled muscle tissue in 1,5h steps between 0 and 6 hours after injection. We quantified labeled and unlabeled phenylalanine both in muscle protein and in the cytosol. This allowed us to critically re-evaluate three different protein synthesis rate (Ks) calculation methodologies that have been developed over the last decades. The calculated values differ by more than 70-fold (0.048 ± 0.021% day-1 up to 3.56 ± 2.16 day-1) between methods. We argue that the Ks calculation including a proportionate ratio of protein synthesis from (unlabeled) free amino acids yields the most realistic Ks values for cold water fish. Eventually, the standardization of the net protein synthesis rate calculation will lead to dependable quantitative representations of organismal stress in response to climate change. |
format |
Dataset |
author |
Krebs, Nina Tebben, Jan Pörtner, Hans-Otto Lannig, Gisela Lucassen, Magnus Mark, Felix Christopher Bock, Christian |
author_facet |
Krebs, Nina Tebben, Jan Pörtner, Hans-Otto Lannig, Gisela Lucassen, Magnus Mark, Felix Christopher Bock, Christian |
author_sort |
Krebs, Nina |
title |
Protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in Antarctic fish |
title_short |
Protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in Antarctic fish |
title_full |
Protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in Antarctic fish |
title_fullStr |
Protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in Antarctic fish |
title_full_unstemmed |
Protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in Antarctic fish |
title_sort |
protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in antarctic fish |
publisher |
PANGAEA |
publishDate |
2022 |
url |
https://doi.pangaea.de/10.1594/PANGAEA.940626 https://doi.org/10.1594/PANGAEA.940626 |
op_coverage |
LATITUDE: 53.533000 * LONGITUDE: 8.580100 |
long_lat |
ENVELOPE(8.580100,8.580100,53.533000,53.533000) |
genre |
Antarc* Antarctic Antarctica |
genre_facet |
Antarc* Antarctic Antarctica |
op_relation |
Krebs, Nina; Bock, Christian; Tebben, Jan; Mark, Felix Christopher; Lucassen, Magnus; Lannig, Gisela; Pörtner, Hans-Otto (2023): Evolutionary Adaptation of Protein Turnover in White Muscle of Stenothermal Antarctic Fish: Elevated Cold Compensation at Reduced Thermal Responsiveness [dataset]. PANGAEA, https://doi.org/10.1594/PANGAEA.963276 https://doi.pangaea.de/10.1594/PANGAEA.940626 https://doi.org/10.1594/PANGAEA.940626 |
op_rights |
UNKNOWN: Licensing unknown: Please contact principal investigator/authors to gain access and request licensing terms Access constraints: access rights needed info:eu-repo/semantics/restrictedAccess |
op_doi |
https://doi.org/10.1594/PANGAEA.94062610.1594/PANGAEA.963276 |
_version_ |
1810494927670345728 |