Absolute quantification of apolipoproteins and associated proteins on human plasma lipoproteins

Zychlinski, Anne von; Williams, Michael J.A.; McCormick, Sally P.A.; Kleffmann, Torsten

doi:10.1016/j.jprot.2014.04.030

Cited by 58 publications

(57 citation statements)

References 38 publications

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“…2A ). Our analysis confi rmed the presence of classical apolipoproteins, complement factors, fi brinogen, serpins, LCAT, and serum paraoxonase/arylesterase 1 (PON1), all of which were previously reported using other HDL isolation methods ( 35,36 ). The protein-specifi c sum-normalized spectral counts (per participant) estimated the relative proportion of protein abundances across the fi ve native gel size fractions.…”

Section: The Hdl Size Fraction Subproteomessupporting

confidence: 76%

“…The input compartment is the plasma amino acid precursor pool (D3-Leu tracer enrichment in plasma) expressed as a forcing function that drives the appearance of plasma D3-Leu tracer in the model. Each participant's D3-Leu tracer by guest, on www.jlr.org that contain low levels of apoA-I (36)(37)(38). Group II represents the proteins that predominate in ␣ 1 and ␣ 2 size HDL and contains 11 proteins including apoE, apoM, apoL-I, apoC-IV, cholesteryl ester transfer protein (CETP), and phospholipid transfer protein (PLTP).…”

Section: Multicompartmental Modelingmentioning

confidence: 99%

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Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring

Singh

Andraski

Pieper

et al. 2016

Journal of Lipid Research

130

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This article is available online at http://www.jlr.org usually determines isotope enrichment by measuring the derivatized forms of D0 and trideuterated leucine (D3-Leu) ( 2, 3 ), a method with high cost and low sensitivity and specifi city. Recently, proteomics-based triple quadrupole multiple reaction monitoring (MRM) permitted a more practical and highly specifi c multipeptide approach to in vivo kinetic studies ( 4, 5 ). However, MRM relies on low-resolution readouts (unit mass resolution) that do not readily permit precise quantifi cation of tracer enrichment that is lower than 1%, which is common in apolipoprotein kinetics ( 5, 6 ). Factors contributing to low precision include interference by not only the sister isotope 13C15N M3 ion but also background ions. In this study, we aim to extend further the scope of in vivo kinetics by exploiting the recently developed highresolution/accurate mass parallel reaction monitoring (HR/AM-PRM) method performed on the quadrupole Orbitrap mass spectrometer ( 7,8 ). The HR/AM fragment ion scan feature has the potential to measure D3-Leu enrichment between 0.03% and 1.0%, a low incorporation range that is a consequence of a bolus-administered tracer, useful in revealing tracer-tracee relationships. (Nagoya, Japan; M.A.) and the National Institutes of Health [ R01HL107550 (M.A.); UL1 RR 025758-01 ; and R01HL095964 (F.M.S.)]. Abstract Endogenous labeling with stable isotopes is used

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Section: The Hdl Size Fraction Subproteomessupporting

confidence: 76%

Section: Multicompartmental Modelingmentioning

confidence: 99%

Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring

Singh

Andraski

Pieper

et al. 2016

Journal of Lipid Research

130

View full text Add to dashboard Cite

show abstract

“…Following the termination of the centrifuge run, fractions were collected and the absorbance at 280 nm was recorded. The amount recovered in each fraction was normalized to the total absorbance for all the recorded fractions ( 22,(32)(33)(34)(35), this may exacerbate the alteration of their recovered HDL; as in this report, we demonstrate that centrifugation of HDL at rotor speeds in excess of 30,000 rpm results in alteration to the recovered particle. Minimally altered HDL can be recovered from plasma samples using a reduced rotor speed of 15,000 rpm and a KBr-containing gradient in a similar time frame as the traditional ultracentrifuge-based isolation.…”

Section: Loss Of Protein From Hdlsupporting

confidence: 62%

Excessive centrifugal fields damage high density lipoprotein [S]

Munroe

Phillips

Schumaker

2015

Journal of Lipid Research

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This article is available online at http://www.jlr.org with centrifugation for 38 h to isolate HDL using a 40.3 or Ti70 rotor at у 40,000 rpm ( 4, 5 ). It was also John Gofman who proposed that elevated levels of lipoproteins cause atherosclerosis, the deposition of lipid in arteries and blood vessels that eventually leads to the premature deaths of so many people in the developed world ( 6 ). It was noticed that HDL could reverse this trend ( 7-9 ), which spurred an intense research focus on the mechanism of HDL as an atheroprotective particle ( 10-15 ), as well as identifying its other functions (16)(17)(18)(19)(20).Few reports have attempted to characterize the stability of lipoproteins in a centrifugal fi eld. Kunitake and Kane ( 21 ) studied human lipoproteins and found that apoAI was lost during each spin until one-third of the apoAI was lost with fi ve spins. Additionally, Ståhlman et al. ( 22 ) demonstrated that using reduced ionic strength solutions allowed for greater recovery of exchangeable apolipoproteins. Increased ionic strength solutions were found to improve the retention of apoAI on the HDL ( 21 ), whereas this decreased the retention of the exchangeable apolipoproteins ( 22 ). Thus, for different apolipoproteins, their mechanism of dissociation from HDL differs. The assembly of HDL is hinted at by less disruptive characterization methods, such as nondenaturing gradient gel electrophoresis. Li et al. ( 23 ) discovered that native gel electrophoresis produced a series of 14 distinct HDL peaks, which they labeled in the order in which they appeared, although most individuals only have about six. LeBoeuf et al. ( 24 ), characterized mouse lipoproteins from a variety of inbred mouse strains and found that mouse lipoproteins were roughly similar in size, although there were differences in the electrophoretic position of the major peak. Thus, depending upon the method of isolation, the pattern of lipoproteins recovered differs.As there are many contemporary publications that utilize high ultracentrifuge rotor speeds, from 65,000 to 120,000 rpm, in order to reduce the time frame needed to isolate lipoproteins ( 25-31 ), and many characterizations of HDL utilize material which was purifi ed via ultracentrifugation Abstract HDL is typically isolated ultracentrifugally at 40,000 rpm or greater, however, such high centrifugal forces are responsible for altering the recovered HDL particle. We demonstrate that this damage to HDL begins at approximately 30,000 rpm and the magnitude of loss increases in a rotor speed-dependent manner. The HDL is affected by elevated ultracentrifugal fi elds resulting in a lower particle density due to the shedding of associated proteins. To circumvent the alteration of the recovered HDL, we utilize a KBr-containing density gradient and a lowered rotor speed of 15,000 rpm to separate the lipoproteins using a single 96 h centrifugation step. This recovers the HDL at two density ranges; the bulk of the material has a density of about 1.115 g/ml, while lessor amounts of material ...

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“…Peptide standards were used to quantify 17 apolipoproteins (one standard per protein) in Lp(a), VLDL, LDL and HDL, isolated from healthy individuals by density centrifugation followed by size exclusion chromatography [16]. The relative distributions of the apolipoproteins within and among the lipoproteins varied; for example, after apoA-I, apoA-II and apoC-II were the second and third most abundant proteins on HDL and after apoB, apoC-II and apoC-III on VLDL and interestingly, apoA-I and apoC-II on LDL [16].…”

Section: Unbiased Proteomics Applied To Hdl Biologymentioning

confidence: 99%

Unbiased and targeted mass spectrometry for the HDL proteome

Singh

Aikawa

2017

Current Opinion in Lipidology

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Purpose of review Mass spectrometry (MS) is an ever evolving technology that is equipped with a variety of tools for protein research. Some lipoprotein studies, especially those pertaining to HDL biology, have been exploiting the versatility of MS to understand HDL function through its proteome. Despite the role of MS in advancing research as a whole however, the technology remains obscure to those without hands on experience, but still wishing to understand it. In this review, we walk the reader through the co-evolution of common MS workflows and HDL research, starting from the basic unbiased MS methods used to profile the HDL proteome to the most recent targeted methods that have enabled an unprecedented view of HDL metabolism. Recent findings Unbiased global proteomics have demonstrated that the HDL proteome is organized into sub-groups across the HDL size fractions providing further evidence that HDL functional heterogeneity is in part governed by its varying protein constituents. Parallel reaction monitoring, a novel targeted MS method, was used to monitor the metabolism of HDL apolipoproteins in humans and revealed that apolipoproteins contained within the same HDL size fraction exhibit diverse metabolic properties. Summary MS provides a variety of tools and strategies to facilitate understanding, through its proteins, the complex biology of HDL.

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Absolute quantification of apolipoproteins and associated proteins on human plasma lipoproteins

Cited by 58 publications

References 38 publications

Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring

Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring

Excessive centrifugal fields damage high density lipoprotein [S]

Unbiased and targeted mass spectrometry for the HDL proteome

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