scholarly journals High-Abundance Polypeptides of the Human Plasma Proteome Comprising the Top 4 Logs of Polypeptide Abundance

2008 ◽  
Vol 54 (10) ◽  
pp. 1608-1616 ◽  
Author(s):  
Glen L Hortin ◽  
Denis Sviridov ◽  
N Leigh Anderson
Keyword(s):  
Plasma Proteins ◽  
Clinical Laboratory ◽  
Plasma Concentrations ◽  
High Abundance ◽  
Molar Basis ◽  
Protein Molecules ◽  
Polypeptide Chains ◽  
Peptide Segments ◽  
Human Plasma Proteome ◽  
Individual Peptide

Abstract Background: Plasma contains thousands of proteins, but a small number of these proteins comprise the majority of protein molecules and mass. Content: We surveyed proteomic studies to identify candidates for high-abundance polypeptide chains. We searched the literature for information on the plasma concentrations of the most abundant components in healthy adults and for the molecular mass of the mature polypeptide chains in plasma. Because proteomic studies usually dissociate proteins into polypeptide chains or detect short peptide segments of proteins, we summarized data on individual peptide chains for proteins containing multiple subunits or polypeptides. We collected data on about 150 of the most abundant polypeptides in plasma. The abundant polypeptides span approximately the top 4 logs of concentration in plasma, from 650 to 0.06 μmol/L on a molar basis or from about 50 000 to 1 mg/L mass abundance. Conclusions: Data on the concentrations of the high-abundance peptide chains in plasma assist in understanding the composition of plasma and potential approaches for clinical laboratory or proteomic analysis of plasma proteins. Development of more extensive databases regarding the plasma concentrations of proteins in health and diseases would promote diagnostic and proteomic advances.

scholarly journals Analysis of free and protein-bound nitrotyrosine in human plasma by a gas chromatography/mass spectrometry method that avoids nitration artifacts

2000 ◽  
Vol 345 (3) ◽  
pp. 453-458 ◽  
Author(s):  
Matthew T. FROST ◽  
Barry HALLIWELL ◽  
Kevin P. MOORE
Keyword(s):  
Reactive Nitrogen Species ◽  
Plasma Proteins ◽  
Biological Fluids ◽  
Plasma Concentrations ◽  
Normal Subjects ◽  
Gas Chromatography Mass Spectrometry ◽  
Highly Sensitive ◽  
Acidic Conditions ◽  
First Time

Measurement of nitrotyrosine in biological fluids and tissues is increasingly being used to monitor the production of reactive nitrogen species in vivo. The detection of nitrotyrosine in vivo has been reported with the use of a variety of methods including immunoassay, HPLC and GLC/MS. The validity of HPLC and immunoassays have been questioned with regard to their selectivity and sensitivity limits. In principle, the measurement of nitrotyrosine by GLC/MS permits a highly specific, highly sensitive and fully quantitative assay. The nitration of tyrosine under acidic conditions in the presence of nitrite is well documented. Derivatization for the full quantification of nitrotyrosine by using GLC/MS can lead to the artifactual nitration of tyrosine if performed under acidic conditions in the presence of nitrite. We describe a novel alkaline method for the hydrolysis and derivatization of nitrotyrosine and tyrosine, and demonstrate its applicability to the measurement of plasma concentrations of both free and protein-bound nitrotyrosine and tyrosine. A detection limit of 1 pg for nitrotyrosine and 100 pg for tyrosine has been achieved. Our method allows, for the first time, the analysis of free and protein-bound nitrotyrosine and tyrosine in biological samples. The plasma concentrations (means±S.E.M.) of free tyrosine and nitrotyrosine in eight normal subjects were 12±0.6 μg/ml and 14±0.7 ng/ml respectively. Plasma proteins contained tyrosine and nitrotyrosine at 60.7±1.7 μg/mg and 2.7±0.4 ng/mg respectively.

Comparison of the feedback effect of magnesium and calcium on parathyroid hormone secretion in man

1987 ◽  
Vol 113 (1) ◽  
pp. 117-122 ◽  
Author(s):  
O. Ferment ◽  
P. E. Garnier ◽  
Y. Touitou
Keyword(s):  
Parathyroid Hormone ◽  
Urinary Excretion ◽  
Hormone Secretion ◽  
Plasma Concentrations ◽  
Magnesium Salt ◽  
Saline Injection ◽  
Molar Basis ◽  
Magnesium Salts ◽  
High Doses

ABSTRACT Administration of high doses of magnesium is known to produce a decrease in parathyroid hormone (PTH) secretion in human patients but the effect of magnesium on the secretion of PTH in healthy man is not known. We have looked at the effect of a relatively moderate i.v. dose of magnesium (7·08 mmol) in seven healthy men. In addition and for comparison the effect of calcium (4·25 mmol) was studied. Two magnesium salts were considered, magnesium sulphate (MgSO4) and magnesium pyrrolidone carboxylate (MgPC). Four i.v. injections were given at 08.00 h (MgPC, NaCl (control), MgSO4 and Ca gluconate), with an interval of 1 week between each injection. Whatever the magnesium salt the variations in plasma concentrations of magnesium were the same whereas no change in erythrocyte magnesium was observed. Plasma concentration of C-terminal PTH did not show significant variations after MgPC or saline injection. Both MgSO4 and Ca gluconate produced a statistically significant 30% decrease in plasma PTH levels 45 min after the injection. The effect was more sustained with calcium (2 h) than with magnesium (45 min). The urinary excretion of magnesium was significantly higher after injection of MgSO4 than after MgPC. These results suggest (1) that magnesium was, on a molar basis, less potent than calcium in regulating PTH secretion in vivo, (2) that the nature of the magnesium salt used must be kept in mind for the interpretation of the effect of magnesium on PTH secretion in vivo and (3) that the decrease in plasma PTH can partly explain the larger urinary excretion of magnesium after MgSO4 than after MgPC. J. Endocr. (1987) 113, 117–122

scholarly journals Establishing an optimized method for the separation of low and high abundance blood plasma proteins

2020 ◽  
Vol 2 ◽  
pp. e6
Author(s):  
Henian Yang ◽  
Guijie Wang ◽  
Tiantian Zhang ◽  
John H. Beattie ◽  
Shaobo Zhou
Keyword(s):  
Blood Plasma ◽  
Plasma Proteins ◽  
Protein Concentration ◽  
Protein Precipitation ◽  
High Abundance ◽  
Good Reproducibility ◽  
Average Ratio ◽  
Optimal Separation ◽  
Elution Buffer ◽  
Blood Plasma Proteins

The study tested the efficiency and reproducibility of a method for optimal separation of low and high abundant proteins in blood plasma. Firstly, three methods for the separation and concentration of eluted (E: low abundance), or bound (B: high abundance) proteins were investigated: TCA protein precipitation, the ReadyPrep™ 2-D cleanup Kit and Vivaspin Turbo 4, 5 kDa ultrafiltration units. Secondly, the efficiency and reproducibility of a Seppro column or a ProteoExtract Albumin/IgG column were assessed by quantification of E and B proteins. Thirdly, the efficiency of two elution buffers, containing either 25% or 10% glycerol for elution of the bound protein, was assessed by measuring the remaining eluted volume and the final protein concentration. Compared to the samples treated with TCA protein precipitation and the ReadyPrep™ 2-D cleanup Kit, the E and B proteins concentrated by the Vivaspin4, 5 kDa ultrafiltration unit were separated well in both 1-D and 2-D gels. The depletion efficiency of abundant protein in the Seppro column was reduced after 15 cycles of sample processing and regeneration and the average ratio of E/(B + E) × 100% was 37 ± 11(%) with a poor sample reproducibility as shown by a high coefficient of variation (CV = 30%). However, when the ProteoExtract Albumin/IgG column was used, the ratio of E/(B + E) × 100% was 43 ± 3.1% (n = 6) and its CV was 7.1%, showing good reproducibility. Furthermore, the elution buffer containing 10% (w/v) glycerol increased the rate of B protein elution from the ProteoExtract Albumin/IgG column, and an appropriate protein concentration (3.5 µg/µl) for a 2-D gel assay could also be obtained when it was concentrated with Vivaspin Turbo 4, 5 kDa ultrafiltration unit. In conclusion, the ProteoExtract Albumin/IgG column shows good reproducibility of preparation of low and high abundance blood plasma proteins when using the elution buffer containing 10% (w/v) glycerol. The optimized method of preparation of low/high abundance plasma proteins was when plasma was eluted through a ProteoExtract Albumin/IgG removal column, the column was further washed with elution buffer containing 10% glycerol. The first and second elution containing the low and high abundance plasma proteins, respectively, were further concentrated using Vivaspin® Turbo 4, 5 kDa ultrafiltration units for 1 or 2-D gel electrophoresis.

scholarly journals The early changes in retinol-binding protein and prealbumin concentrations in plasma of protein–energy malnourished children after treatment with retinol and an improved diet

1980 ◽  
Vol 43 (3) ◽  
pp. 393-402 ◽  
Author(s):  
Suzanne Large ◽  
G. Neal ◽  
J. Glover ◽  
O. Thanangkul ◽  
R. E. Olson
Keyword(s):  
Body Weight ◽  
Vitamin A ◽  
Nutritional Status ◽  
Plasma Proteins ◽  
Binding Protein ◽  
Vitamin A Deficiency ◽  
Plasma Concentrations ◽  
Retinol Binding Protein ◽  
Mean Values ◽  
The Mean

1. Changes in total retinol-binding protein (RBP), the holoprotein (holoRBP) and prealbumin (PA) concentrations have been monitored in plasma of thirty protein- and vitamin A-deficient preschool children from within a few hours up to 7 weeks after treatment with retinol and a good-quality protein diet.2. The children were classified into groups according to nutritional status as having either kwashiorkor, marasmus-kwashiorkor or marasmus, and given formula diets whose protein and energy contents increased stepwise from 1 g and 105 kJ/kg body-weight respectively up to 4 g and 733 kJ/kg body-weight after 4 weeks. Retinol was administered in the forms of retinyl palmitate either orally or intramuscularly.3. PA and total RBP were determined by electroimmunoassay procedures and the holoRBP by its fluorescence after separation from other plasma proteins.4. RBP in plasma of the vitamin A-deficient child is largely denatured and incapable of binding administered retinol, which must first be taken up by the liver before native holoRBP is released. An increased pool of native apoprotein accumulates in the liver during vitamin A deficiency which is released into plasma quickly after retinol uptake to form peak concentrations of total and holoRBP approximately 3 h after dosing intramuscularly and 6 h orally.5. The accumulated pool of RBP was highest in livers from the marasmus group and lowest in those from the kwashiorkor group, reflecting their relative capacities to synthesize plasma proteins.6. The mean plasma concentrations of total and holoRBP for the various groups were minimal 24–48 h after dosing with retinol and then improved almost linearly over the following week.7. Mean plasma PA concentrations of the various groups on admission were also in order of the severity of their malnutrition. There was little or no change in this protein concentration over the first 24 h after dosing with retinol, but thereafter the mean values rose almost linearly over 2 weeks. Albumin on the other hand changed little during the first week. The results show that PA is the more sensitive measurement of protein nutritional status.

Eleven Plasma Proteins as Indicators of Protein Nutritional Status in Very Low Birth Weight Infants

PEDIATRICS ◽  
1990 ◽  
Vol 86 (6) ◽  
pp. 916-921 ◽  
Author(s):  
Staffan K. T. Polberger ◽  
Göran A. Fex ◽  
Irene E. Axelsson ◽  
Niels C. R. Räihä
Keyword(s):  
Birth Weight ◽  
Low Birth Weight ◽  
Nutritional Status ◽  
Protein Intake ◽  
Plasma Proteins ◽  
Very Low Birth Weight ◽  
Binding Protein ◽  
Plasma Concentrations ◽  
Retinol Binding Protein ◽  
Low Birth Weight Infants

Concentrations of 11 plasma proteins were measured in 28 healthy, growing, very low birth weight, appropriate-for-gestational-age infants fed varying levels of human milk protein intake (range 1.7 to 3.9 g/kg per day). Significant positive correlations were found between ween mean protein intake and concentrations of 7 of the plasma proteins studied (transthyretin, retinol-binding protein, and transferrin: P < .001; vitamin D-binding protein and apolipoprotein B: P < .01; albumin and apolipoprotein A I: P < .05). A weak negative correlation with mean protein intake was seen for the plasma level of orosomucoid, whereas no significant correlations were found for the plasma concentrations of fibronectin and α1-antichymotrypsin. Protein intake, not energy intake, constituted the main contribution to the changes in the concentrations of transthyretin, retinol-binding protein, and transferrin. The levels of plasma transthyretin and transferrin were also strongly correlated with weight and length growth of the infants during the study as well as with other indicators of protein nutritional status such as preprandial concentrations of plasma amino acids and serum and urine urea. These data indicate that of the 11 plasma proteins studied, transthyretin, transferrin, and retinol-binding protein are the most suitable to evaluate protein nutritional status in very low birth weight infants.

Association of Epileptic and Non-epileptic Seizures and Changes in Circulating Plasma Proteins Linked to Neuroinflammation

Neurology ◽  
2021 ◽  
pp. 10.1212/WNL.0000000000011552
Author(s):  
John Gledhill ◽  
Elizabeth Brand ◽  
John Pollard ◽  
Richard St. Clair ◽  
Todd Wallach ◽  
...  
Keyword(s):  
Risk Factors ◽  
Diagnostic Test ◽  
Epileptic Seizure ◽  
Plasma Proteins ◽  
Sandwich Elisa ◽  
Plasma Concentrations ◽  
Diagnostic Algorithm ◽  
Epileptic Seizures ◽  
Cost Effective ◽  
Class Iii

Objective:To develop a diagnostic test that stratifies epileptic seizure (ES) from psychogenic non-epileptic seizure (PNES) by developing a multimodal algorithm that integrates plasma concentrations of selected immune response associated proteins and patient clinical risk factors for seizure.Methods:Daily blood samples were collected from patients evaluated in the epilepsy monitoring unit (EMU) within 24 hours after EEG confirmed ES or PNES and plasma was isolated. Levels of 51 candidate plasma proteins were quantified using an automated, multiplexed, sandwich ELISA and then integrated and analyzed using our diagnostic algorithm.Results:A 51 protein multiplexed ELISA panel was used to determine the plasma concentrations of ES patients, PNES patients, and healthy controls. A combination of protein concentrations, TRAIL, ICAM-1, MCP-2 and TNF-R1 provided a probability that a patient recently experienced a seizure with TRAIL and ICAM-1 higher in PNES than ES, and MCP-2 and TNF-R1 higher in ES than PNES. The diagnostic algorithm yielded an AUC of 0.94±0.07, sensitivity of 82.6% (95% CI:62.9-93.0) and specificity of 91.6% (95% CI:74.2-97.7). Further, expanding the diagnostic algorithm to include previously identified PNES risk factors enhanced diagnostic performance with AUC of 0.97±0.05, sensitivity of 91.3%; (95% CI:73.2-97.6), and specificity of 95.8%; (95% CI:79.8-99.3).Conclusions:These four plasma proteins could provide a rapid, cost-effective, and accurate blood-based diagnostic test to confirm recent ES or PNES.Classification of Evidence:This study provides Class III evidence that variable levels of four plasma proteins, when analyzed by a diagnostic algorithm, can distinguish PNES from ES with sensitivity of 82.6% and specificity of 91.6%.

Safety and Pharmacokinetics of Standardized Extract of Centella asiatica (ECa 233) Capsules in Healthy Thai Volunteers: A Phase 1 Clinical Study

Planta Medica ◽  
2019 ◽  
Vol 85 (06) ◽  
pp. 483-490 ◽  
Author(s):  
Phanit Songvut ◽  
Pajaree Chariyavilaskul ◽  
Mayuree Tantisira ◽  
Phisit Khemawoot
Keyword(s):  
Clinical Laboratory ◽  
Phase 1 ◽  
Plasma Concentrations ◽  
Centella Asiatica ◽  
Fold Increase ◽  
Repeated Dose ◽  
Pharmacokinetic Parameters ◽  
Maximum Plasma ◽  
Asiatic Acid ◽  
Active Metabolites

AbstractThe aim of this study was to investigate the safety and pharmacokinetic profiles of a newly developed, standardized extract of Centella asiatica (ECa 233) capsule in healthy Thai volunteers. This study was designed as an open-labeled, 2-sequence dosage, single- and repeated-dose study investigated under fasting conditions. Plasma concentrations of the parent compounds and their relative acid metabolites were measured and pharmacokinetic parameters were calculated using noncompartmental analysis. Tolerability was assessed based on physical examinations, monitoring of vital signs, clinical laboratory tests, and any observed adverse events. A key finding of this study was that the pharmacokinetics of ECa 233 in healthy volunteers did not correspond with its pharmacokinetics in animal studies. As indicated in human pharmacokinetic parameters, maximum plasma concentration and area under the curve of the parent compounds (madecassoside and asiaticoside) were very low, while their respective metabolites (madecassic acid and asiatic acid) demonstrated higher values. Based on the pharmacokinetic results observed in the dose comparison, accumulation of active metabolites after repeated dose is highly suggestive. In addition, the asiatic acid profile showed 2-fold increase in Cmax and AUC(0–t) after increasing dose from 250 to 500 mg of ECa 233. Lastly, the safety and tolerability evaluation illustrated that single and multiple doses in both 250 and 500 mg oral administration of ECa 233 were well tolerated, and none of the volunteers discontinued their participation due to adverse effects during the study.

The Role of β2-Glycoprotein I in the Clearance of Platelet Microvesicles.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 145-145
Author(s):  
Hanan Abdel-Monem ◽  
Swapan Kumar Dasgupta ◽  
Anhquyen Le ◽  
Anthony Prakasam ◽  
Perumal Thiagarajan
Keyword(s):  
Plasma Proteins ◽  
Plasma Concentrations ◽  
Major Protein ◽  
Maximal Effect ◽  
Dependent Manner ◽  
Anionic Phospholipid ◽  
Concentration Dependent Manner ◽  
Glycoprotein I

Abstract Abstract 145 The physiological function of β2-glycoprotein I is unclear and several studies suggest a role in the clearance of anionic phospholipid containing membranes. Anionic phospholipid containing liposomes are cleared rapidly from the circulation by the reticuloendothelial cells. In rats, uptake of liposomes by Kupffer cells requires that the liposomes bind to plasma proteins. In mice, the clearance of liposomes from the circulation is related to their ability to interact with plasma proteins. β2-glycoprotein I was identified as a major protein associated with rapid clearance of liposomes and pretreating the mice with antiβ2- glycoprotein I antibodies was found to significantly increase the half-life of the liposome. In vitro, β2-glycoprotein I was also shown to promote the phagocytosis of phosphatidylserine containing liposomes and apoptotic tumor cells. In conditions associated with increased microvesicles generation such as disseminated intravascular coagulation, plasma levels of β;2-glycoprotein I was reduced presumably due to its consumption. Antibodies to β2 glycoprotein I are frequently seen in patients with systemic lupus erythematosus and at times, in otherwise normal individuals. A subset of these antibodies prevents the assembly of the prothrombinase and the tenase complexes on phospholipid membrane, leading to the lupus anticoagulant effect. The presence of these antibodies is clinically very significant, as individuals harboring these antibodies are at risk for thromboembolic manifestations. We studied the role of β-glycoprotein I in the clearance of procoagulant platelet microvesicles and the effect of the auto antibodies in the phagocytosis of platelet microvesicles. We labeled β2-glycoprotein I with BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene)-hydrazide and β2-glycoprotein I incorporated 1.8 mole of BODIPY /mole. Labeling of β2-glycoprotein I with BODIPY did not change the binding efficacy of β2-glycoprotein I to cardiolipin as determined by Elisa assay. Binding of BODIPY-β2-glycoprotein I to platelet microvesicles was analyzed by flow cytometry. BODIPY- β2-glycoprotein I bound to phosphatidylserine-expressing platelet microvesicles in a concentration-dependent manner. Binding was inhibited by 50 fold molar excess of unlabeled β2-glycoprotein I, annexin A5 and the phosphatidylserine-binding C1C2 fragment of lactadherin. β2-glycoprotein I also promoted the phagocytosis of platelet microvesicles by THP-1 derived macrophages in vitro at physiological plasma concentrations with a half maximal effect at ∼10 ug/ml. β2-glycoprotein I-mediated phagocytosis was inhibited by annexin V and the C1C2 fragment of lactadherin. Furthermore, immunoaffinity purified β2-glycoprotein I-dependent antiphospholipid antibodies from 5 patients inhibited the phagocytosis in a concentration dependent manner. These studies suggest β2-glycoprotein I binding to phosphatidylserine-expressing procoagulant platelet microvesicles promotes their clearance by macrophages and autoantibodies to β2-glycoprotein I inhibit the process. The predictive value of antiβ-2 glycoprotein I for thrombosis is highly variable but the correlation is stronger in patients with lupus. In lupus, there is impaired clearance of procoagulant apoptotic cells. β2-glycoprotein I may have a significant role in their clearance and antibodies to β2-glycoprotein I may causally related to the thrombosis in these patients by inhibiting the clearance. Disclosures: No relevant conflicts of interest to declare.

Structure and Mechanical Properties of Rubberlike Proteins in Animals

1987 ◽  
Vol 60 (3) ◽  
pp. 417-438 ◽  
Author(s):  
John M. Gosline
Keyword(s):  
Amorphous Polymer ◽  
Polymer Networks ◽  
Random Coil ◽  
Future Research ◽  
Random Networks ◽  
Glass Transitions ◽  
Mechanical State ◽  
Protein Molecules ◽  
Polypeptide Chains ◽  
Covalent Crosslinks

Abstract Polymer networks formed from protein molecules that adopt kinetically-free, random-coil conformations are found in many animals, where they play a number of important roles. The 5 rubberlike proteins isolated and studied to date indicate that animal rubbers, like their synthetic counterparts, contain random networks which are usually stabilized by covalent crosslinks. Long-range elasticity in rubberlike proteins is based on changes in the conformational entropy of random-coil molecules. Further, these protein networks show viscoelastic glass transitions similar to all other amorphous polymer networks. Future research on protein sequences should increase our understanding of how polypeptide chains can function as random-coil molecules, and studies into the mechanical state of elastin in arterial tissues may provide important clues about the mechanisms of some forms of human disease.

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