Dataset Information

Serum proteomics in cats with cardiomyopathy and congestive heart failure

ABSTRACT: In this study, we aimed to identify novel serum biomarkers in feline primary cardiomyopathies with congestive heart failure, which can help to understand disease pathogenesis and assist the disease diagnosis, prognosis and management. The study included 15 cats diagnosed with CHF and CM, 5 cats with asymptomatic CM and 15 healthy cats.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Felis Catus (cat) (felis Silvestris Catus)

TISSUE(S): Blood Serum

DISEASE(S): Cardiovascular System Disease

SUBMITTER: Anita Horvatic

LAB HEAD: Peter David Eckersall

PROVIDER: PXD017761 | Pride | 2020-07-19

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20170914_Tina_Six1.mgf	Mgf
	20170914_Tina_Six1.raw	Raw
	20170914_Tina_six1_7_PSMs.txt	Txt
	20170914_Tina_six1_7_PeptideGroups.txt	Txt
	20170914_Tina_six1_7_ProteinGroups.txt	Txt

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Publications

Novel biomarkers in cats with congestive heart failure due to primary cardiomyopathy.

Liu Mengmeng M Eckersall P David PD Mrljak Vladimir V Horvatić Anita A Guillemin Nicolas N Galan Asier A Köster Liza L French Anne A

Journal of proteomics 20200709

The pathogenesis of feline cardiomyopathy and congestive heart failure (CHF) requires further understanding. In this study, we assessed serum proteome change in feline CHF, aiming to identify novel biomarker for both research and clinical use. The study comprised 15 cats in CHF, 5 cats in preclinical cardiomyopathy and 15 cats as healthy controls. Serum proteome profiles were obtained by tandem mass tag labelling followed by mass spectrometry. Protein concentrations in CHF cats were compared wit ...[more]

PMID: 32652222

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Project description:Approximately 2x106 CL1-5 and H23 cells were seeded on 10-cm dishes with RPMI-1640 medium containing 10% fetal bovine serum. After the cells covered 80% of the dish, cells were serum-starved for 24 h in RPMI-1640 medium. CL1-5 and H23 cell lysates were harvested, trypsinized and analyzed by using LTQ-Orbitrap XL and LTQ-Orbitrap Discovery, respectively. Halobacterium salinarum NRC-1 (ATCC700922) starter culture was prepared by inoculating a single colony in 5 mL of Halobacterium medium containing trace metals (CM+) and grown at 37 °C with shaking at 225 rpm for one week. An aliquot of 300 μL of the culture was spread on 15-cm diameter CM+ plates containing 2% of agarose. After 7~10 days of incubation at 37 °C, the GVs from cells on 10 plates were harvested using the centrifugation facilitated flotation method as previously described except the GVs were washed for a total of 5 times instead of the suggested 3 times to remove nonspecific bound proteins.(29) After collecting the gas vesicles in the primary cell lysate, the cell membrane and residual GV were removed by ultracentrifugation at 53 000× g, 4 °C for 16 h twice to obtain the GV-depleted lysate (GVD). GVD lysate was harvested, trypsinized and analyzed by using LC-MS/MS ( LTQ-Orbitrap XL). The H. salinarum strain NRC-1 was grown in complex medium (CM+) at 37 oC with shaking (200 rpm) to an OD600 of 0.5 (early exponential phase. The cultures were then subjected to an additional hour of incubation at 37 oC for preparation of proteins. The total lysate was harvested, trypsinized and analyzed by using LC-MS/MS (LTQ-Orbitrap Discovery). All MS raw data in Thermo XCalibur (version 2.0.7) binary format were converted to the mzXML open data format using a modified version of the ReAdW program. SEQUEST (version 27) was applied to search the MS/MS spectra against the forward and reverse (decoy) protein sequences of H. salinarum (2,427 proteins; EMBL-EBI Integr8, www.ebi.ac.uk/integr8/EBI-Integr8-HomePage.do) or human (38,425 proteins) plus bovine (22,863 proteins) (GenBank reference sequences, www.ncbi.nlm.nih.gov). SEQUEST results were further processed using the Trans Proteomic Pipeline.