Citation:

PANGAEA.940626

Name: Protein synthesis rates derived from incorporation of non-radioactively labeled phenylalanine in Antarctic fish
Published: 2022-02-03
Keywords: 13C-labeling; Antarctica; cold adaptation; Energy budget; physiology

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Replaced by:

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

Abstract:

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.

Keyword(s):

13C-labeling; Antarctica; cold adaptation; Energy budget; physiology

Coverage:

Latitude: 53.533000 * Longitude: 8.580100

Event(s):

AWI_lab * Latitude: 53.533000 * Longitude: 8.580100 * Location: Bremerhaven, Germany * Method/Device: Experiment (EXP)

Comment:

** DO NOT USE ** please refer to the dataset given in the "Replaced by" reference

Parameter(s):

#	Name	Short Name	Unit	Principal Investigator	Method/Device	Comment
1	Sample code/label	Sample label		Krebs, Nina
2	Sample code/label 2	Sample label 2		Krebs, Nina
3	Subsample ID	Subsample ID		Krebs, Nina
4	Time in hours	Time	h	Krebs, Nina		Injection Time
5	Pachycara brachycephalum, standard length	P. brachycephalum SL	cm	Krebs, Nina	Tape measure
6	Pachycara brachycephalum, mass	P. brachycephalum m	g	Krebs, Nina	Scale
7	Pachycara brachycephalum, liver, mass	P. brachycephalum liver m	g	Krebs, Nina	Scale
8	Hepato-somatic index	HSI	%	Krebs, Nina	Calculated	liver weight / weight *100
9	Sample, wet mass	Sample wm	mg	Krebs, Nina	Scale
10	Proteins, dry mass	Protein dm	mg	Krebs, Nina	Scale	dry protein weight during extraction
11	Phenylalanine, unlabelled, in cytosol, area	Phe C12 cytosol	#	Tebben, Jan	Liquid chromatography-high resolution mass spectrometry (LC-HRMS) with Orbitrap
12	Phenylalanine, labelled, in cytosol, area	Phe C13 cytosol	#	Tebben, Jan	Liquid chromatography-high resolution mass spectrometry (LC-HRMS) with Orbitrap
13	Phenylalanine, labelled, in cytosol	Phe C13 cytosol	%	Krebs, Nina
14	Phenylalanine, unlabelled, in protein, area	Phe C12 protein	#	Tebben, Jan	Liquid chromatography-high resolution mass spectrometry (LC-HRMS) with Orbitrap
15	Phenylalanine, labelled, in protein, area	Phe C13 protein	#	Tebben, Jan	Liquid chromatography-high resolution mass spectrometry (LC-HRMS) with Orbitrap
16	Proteins, synthesis rate, per day	Protein synthesis	%/day	Krebs, Nina	Calculated	KsA: (Cytosol_C13_MSMS/Protein_C13_MSMS)(24/IncubationTime)100
17	Proteins, synthesis rate, per day	Protein synthesis	%/day	Krebs, Nina	Calculated	KsB: (KsA(100/Cytosol_C13)100
18	Proteins, synthesis rate, per day	Protein synthesis	%/day	Krebs, Nina	Calculated	KsC: (Protein_C13_MSMS/Protein_C13_MSMS+Protein_C12_MSMS)(100/Cytosol_C13)(24/IncubationTime)*100

License:

Licensing unknown: Please contact principal investigator/authors to gain access and request licensing terms (UNKNOWN)

Status:

Curation Level: Enhanced curation (CurationLevelC)

Size:

1993 data points

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