Evidence map›Paper›PMID 25804408›Full record

ArticleAnalytical biochemistry2015

Quantifying vitamin K-dependent holoprotein compaction caused by differential γ-carboxylation using high-pressure size exclusion chromatography.

Nicholas C Vanderslice, Amanda S Messer, Kanagasabai Vadivel, S Paul Bajaj, Martin Phillips, Mostafa Fatemi, Weijie Xu, William H Velander

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Analytical biochemistry, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed.

  1. Mobile Affinity Sorbent Chromatography.Analytical chemistry · 2018
    Article
  2. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors.

Nicholas C VandersliceProtein Purification and Characterization Laboratories, Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.
Amanda S MesserProtein Purification and Characterization Laboratories, Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA; Protein Science Laboratory, UCLA/Orthopaedic Hospital, Department of Orthopaedic Surgery and Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Kanagasabai VadivelProtein Science Laboratory, UCLA/Orthopaedic Hospital, Department of Orthopaedic Surgery and Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
S Paul BajajProtein Science Laboratory, UCLA/Orthopaedic Hospital, Department of Orthopaedic Surgery and Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Martin PhillipsUCLA-DOE Biochemistry Instrumentation Facility, Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Mostafa FatemiProtein Purification and Characterization Laboratories, Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.
Weijie XuProtein Purification and Characterization Laboratories, Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.
William H VelanderProtein Purification and Characterization Laboratories, Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA. Electronic address: wvelander2@unl.edu.

Funding

cGMP Recombinant FIX for IV &Oral Hemophilia B TherapyR01HL078944 · NHLBI · UNIVERSITY OF NEBRASKA LINCOLN · PI VELANDER, WILLIAM H · 2005 to 2009
$9.6M
NHLBI NIH HHS R01 HL078944PHS HHS R01 JL078944
6 · The paper itself

Abstract

This study uses high-pressure size exclusion chromatography (HPSEC) to quantify divalent metal ion (X(2+))-induced compaction found in vitamin K-dependent (VKD) proteins. Multiple X(2+) binding sites formed by the presence of up to 12 γ-carboxyglutamic acid (Gla) residues are present in plasma-derived FIX (pd-FIX) and recombinant FIX (r-FIX). Analytical ultracentrifugation (AUC) was used to calibrate the Stokes radius (R) measured by HPSEC. A compaction of pd-FIX caused by the filling of Ca(2+) and Mg(2+) binding sites resulted in a 5 to 6% decrease in radius of hydration as observed by HPSEC. The filling of Ca(2+) sites resulted in greater compaction than for Mg(2+) alone where this effect was additive or greater when both ions were present at physiological levels. Less X(2+)-induced compaction was observed in r-FIX with lower Gla content populations, which enabled the separation of biologically active r-FIX species from inactive ones by HPSEC. HPSEC was sensitive to R changes of approximately 0.01nm that enabled the detection of FIX compaction that was likely cooperative in nature between lower avidity X(2+) sites of the Gla domain and higher avidity X(2+) sites of the epidermal growth factor 1 (EGF1)-like domain.

Indexed as

1-Carboxyglutamic AcidBinding SitesCalciumCations, DivalentChromatography, GelFactor IXHumansMagnesiumModels, MolecularProtein ConformationRecombinant ProteinsVitamin K1-Carboxyglutamic AcidCalciumCations, DivalentFactor IXMagnesiumRecombinant ProteinsVitamin KFactor IXHigh-pressure size exclusion chromatographyVitamin K-dependent proteinγ-Carboxylation

Identifiers

PMID25804408
PMCPMC4428943

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.