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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

12 Issues per year


IMPACT FACTOR 2016: 3.432

CiteScore 2016: 2.21

SCImago Journal Rank (SJR) 2016: 1.000
Source Normalized Impact per Paper (SNIP) 2016: 1.112

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1437-4331
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Volume 46, Issue 12

Issues

Accurate assessment and identification of naturally occurring cellular cobalamins

Luciana Hannibal
  • 1Department of Cell Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA and School of Biomedical Sciences, Kent State University, Kent, OH, USA
  • Other articles by this author:
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/ Armend Axhemi / Alla V. Glushchenko / Edward S. Moreira
  • 4Department of Cell Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA and School of Biomedical Sciences, Kent State University, Kent, OH, USA
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/ Nicola E. Brasch
  • 5Department of Cell Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA and Department of Chemistry, Kent State University, Kent, OH, USA
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/ Donald W. Jacobsen
  • 6Department of Cell Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA and Department of Chemistry, Kent State University, Kent, OH, USA and Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine, Case Western Reserve University, Cleveland, OH, USA
  • Other articles by this author:
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Published Online: 2008-10-31 | DOI: https://doi.org/10.1515/CCLM.2008.356

Abstract

Background: Accurate assessment of cobalamin profiles in human serum, cells, and tissues may have clinical diagnostic value. However, non-alkyl forms of cobalamin undergo β-axial ligand exchange reactions during extraction, which leads to inaccurate profiles having little or no diagnostic value.

Methods: Experiments were designed to: 1) assess β-axial ligand exchange chemistry during the extraction and isolation of cobalamins from cultured bovine aortic endothelial cells, human foreskin fibroblasts, and human hepatoma HepG2 cells, and 2) to establish extraction conditions that would provide a more accurate assessment of endogenous forms containing both exchangeable and non-exchangeable β-axial ligands.

Results: The cobalamin profile of cells grown in the presence of [57Co]-cyanocobalamin as a source of vitamin B12 shows that the following derivatives are present: [57Co]-aquacobalamin, [57Co]-glutathionylcobalamin, [57Co]-sulfitocobalamin, [57Co]-cyanocobalamin, [57Co]-adenosylcobalamin, [57Co]-methylcobalamin, as well as other yet unidentified corrinoids. When the extraction is performed in the presence of excess cold aquacobalamin acting as a scavenger cobalamin (i.e., “cold trapping”), the recovery of both [57Co]-glutathionylcobalamin and [57Co]-sulfitocobalamin decreases to low but consistent levels. In contrast, the [57Co]-nitrocobalamin observed in extracts prepared without excess aquacobalamin is undetectable in extracts prepared with cold trapping.

Conclusions: This demonstrates that β-ligand exchange occurs with non-covalently bound β-ligands. The exception to this observation is cyanocobalamin with a non-covalent but non-exchangeable CN group. It is now possible to obtain accurate profiles of cellular cobalamins.

Clin Chem Lab Med 2008;46:1739–46.

Keywords: β-axial ligand exchange; cobalamin; cold trapping; glutathionylcobalamin; vitamin B12

About the article

Corresponding authors: Dr. Donald W. Jacobsen, Dr. Nicola E. Brasch, Department of Cell Biology, NC-10, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA Phone: +1-216-444-8340, Fax: +1-216-444-9404, ,


Received: 2008-07-01

Accepted: 2008-09-11

Published Online: 2008-10-31

Published in Print: 2008-12-01


Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 46, Issue 12, Pages 1739–1746, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2008.356.

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©2008 by Walter de Gruyter Berlin New York. Copyright Clearance Center

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