Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Holzer, M; Kern, S; Birner-Grünberger, R; Curcic, S; Heinemann, A; Marsche, G.
Refined purification strategy for reliable proteomic profiling of HDL2/3: Impact on proteomic complexity.
Sci Rep. 2016; 6(368):38533-38533
Doi: 10.1038/srep38533
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- Führende Autor*innen der Med Uni Graz
- Holzer Michael
- Co-Autor*innen der Med Uni Graz
- Birner-Grünberger Ruth
- Curcic Sanja
- Heinemann Akos
- Marsche Gunther
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- Abstract:
- Proteomics have extended the list of high-density lipoprotein (HDL) associated proteins to about 90. One of the major issues of global protein characterization is establishing specificity of association as opposed to contamination, a fact which has never been addressed for isolated HDL. We have developed a refined purification strategy to isolate HDL by density, followed by purification by size to generate "highly purified" fractions of HDL2/3, which allow the reliable quantification of the HDL proteome for biomarker discovery. Mass spectrometry analysis revealed that the proteome of HDL2/3 is composed of 10-16 different proteins, which is in striking contrast to previous reports. Importantly, proteomic analysis revealed that many proteins which have recently been described to be associated with HDL, including α-1-antitrypsin, α-2-HS-glycoprotein, serotransferrin, apolipoprotein A-IV and others, are not associated with HDL2/3 and are exclusively found in a different molecular weight fraction containing human serum albumin, lipid-poor apolipoprotein A-I and other proteins. Interestingly, proteins found in this lower molecular weight fraction commonly share lipid-binding properties and enrichment of serum with free fatty acids/lysophophatidylcholine led to a significant increase in co-isolation of lipid-binding proteins such as albumin and α-1-antitrypsin. We propose that this refined method might become a standard in proteomic assessment of HDL2/3 making data from clinical cohorts more comparable and reproducible.
- Find related publications in this database (using NLM MeSH Indexing)
- Cross-Linking Reagents - chemistry
- Fatty Acids - blood
- Humans -
- Immunoblotting -
- Lipoproteins, HDL - isolation & purification
- Phospholipases A2 - metabolism
- Phospholipids - blood
- Proteome - metabolism
- Proteomics - methods