Normal bone deposition occurs in mice deficient in Factor XIII-A and Transglutaminase 2

Paul A. Cordell; Laura M. Newell; Kristina F. Standeven; Penelope J. Adamson; Kingsley R. Simpson; Kerrie A. Smith; Christopher L. Jackson; Peter J. Grant; Richard J. Pease

doi:10.1016/j.matbio.2015.02.001

Normal bone deposition occurs in mice deficient in Factor XIII-A and Transglutaminase 2

Paul A. Cordell, Laura M. Newell, Kristina F. Standeven, Penelope J. Adamson, Kingsley R. Simpson, Kerrie A. Smith, Christopher L. Jackson, Peter J. Grant, Richard J. Pease

Research output: Contribution to journal › Article › peer-review

16 Citations (Scopus)

Abstract

Transglutaminase activity has been widely implicated in bone deposition. A predominant role has been proposed for factor (F)XIII-A and a subsidiary role suggested for the homologous protein, transglutaminase 2. Full-length FXIII-A is an 83kDa protransglutaminase that is present both in plasma and also in haematopoietic and connective tissue lineages. Several studies have reported expression in murine cells, including osteocytes, of a 37kDa protein that reacts with the monoclonal anti-FXIII-A antibody AC-1A1. This protein was presumed to be a catalytically active fragment of FXIII-A-83 and to play a major role in bone deposition.

We detected a 37kDa AC-1A1 reactive protein in FXIII-A mRNA negative cell lines and in tissues from FXIII-A^-/- mice. By mass spectrometric sequencing of AC-1A1 immunoprecipitates, we identified this protein as transaldolase-1, and confirmed that recombinant transaldolase-1 is recognised by AC-1A1. We have also shown that bone deposition is normal in FXIII-A^-/-.TG2^-/- double knockout mice, casting doubt on the role of transglutaminases in bone mineralisation. Various studies have used antibody AC-1A1 for immunohistochemistry or immunofluorescence. We observe strong FXIII-A dependent staining in paraffin embedded mouse heart sections, with relatively low background in non-expressing mouse cells. In contrast, FXIII-A independent staining predominates in cultured human cells using a standard immunofluorescence procedure. Immunofluorescence is present in membrane compartments that are expected to lack transaldolase, indicating that other off-target antigens are recognised by AC-1A1.

This has significant implications for studies that have used this approach to define the subcellular trafficking of FXIII-A in osteocytes.

Original language	English
Pages (from-to)	85-96
Number of pages	12
Journal	Matrix Biology
Volume	43
DOIs	https://doi.org/10.1016/j.matbio.2015.02.001
Publication status	Published - 1 Apr 2015
Externally published	Yes

Keywords

Antibodies
Bone development
FXIII-A
Mice
Transaldolase
Transglutaminases

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10.1016/j.matbio.2015.02.001

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@article{97f094baf31c4da195e61fa57cd0b7d8,

title = "Normal bone deposition occurs in mice deficient in Factor XIII-A and Transglutaminase 2",

abstract = "Transglutaminase activity has been widely implicated in bone deposition. A predominant role has been proposed for factor (F)XIII-A and a subsidiary role suggested for the homologous protein, transglutaminase 2. Full-length FXIII-A is an 83kDa protransglutaminase that is present both in plasma and also in haematopoietic and connective tissue lineages. Several studies have reported expression in murine cells, including osteocytes, of a 37kDa protein that reacts with the monoclonal anti-FXIII-A antibody AC-1A1. This protein was presumed to be a catalytically active fragment of FXIII-A-83 and to play a major role in bone deposition. We detected a 37kDa AC-1A1 reactive protein in FXIII-A mRNA negative cell lines and in tissues from FXIII-A-/- mice. By mass spectrometric sequencing of AC-1A1 immunoprecipitates, we identified this protein as transaldolase-1, and confirmed that recombinant transaldolase-1 is recognised by AC-1A1. We have also shown that bone deposition is normal in FXIII-A-/-.TG2-/- double knockout mice, casting doubt on the role of transglutaminases in bone mineralisation. Various studies have used antibody AC-1A1 for immunohistochemistry or immunofluorescence. We observe strong FXIII-A dependent staining in paraffin embedded mouse heart sections, with relatively low background in non-expressing mouse cells. In contrast, FXIII-A independent staining predominates in cultured human cells using a standard immunofluorescence procedure. Immunofluorescence is present in membrane compartments that are expected to lack transaldolase, indicating that other off-target antigens are recognised by AC-1A1. This has significant implications for studies that have used this approach to define the subcellular trafficking of FXIII-A in osteocytes.",

keywords = "Antibodies, Bone development, FXIII-A, Mice, Transaldolase, Transglutaminases",

author = "Cordell, {Paul A.} and Newell, {Laura M.} and Standeven, {Kristina F.} and Adamson, {Penelope J.} and Simpson, {Kingsley R.} and Smith, {Kerrie A.} and Jackson, {Christopher L.} and Grant, {Peter J.} and Pease, {Richard J.}",

note = "Funding Information: We thank the British Heart Foundation for Project Grant support ( PG/03/080/15731 ) and for a non-clinical PhD studentship (FS/08/050/25667) and also the Leeds Multidisciplinary Cardiovascular Research Centrefor a Career Development Grant (to PAC). Publisher Copyright: {\textcopyright} 2015.",

year = "2015",

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doi = "10.1016/j.matbio.2015.02.001",

language = "English",

volume = "43",

pages = "85--96",

journal = "Matrix Biology",

issn = "0945-053X",

publisher = "Elsevier B.V.",

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AU - Cordell, Paul A.

AU - Newell, Laura M.

AU - Standeven, Kristina F.

AU - Adamson, Penelope J.

AU - Simpson, Kingsley R.

AU - Smith, Kerrie A.

AU - Jackson, Christopher L.

AU - Grant, Peter J.

AU - Pease, Richard J.

N1 - Funding Information: We thank the British Heart Foundation for Project Grant support ( PG/03/080/15731 ) and for a non-clinical PhD studentship (FS/08/050/25667) and also the Leeds Multidisciplinary Cardiovascular Research Centrefor a Career Development Grant (to PAC). Publisher Copyright: © 2015.

PY - 2015/4/1

Y1 - 2015/4/1

N2 - Transglutaminase activity has been widely implicated in bone deposition. A predominant role has been proposed for factor (F)XIII-A and a subsidiary role suggested for the homologous protein, transglutaminase 2. Full-length FXIII-A is an 83kDa protransglutaminase that is present both in plasma and also in haematopoietic and connective tissue lineages. Several studies have reported expression in murine cells, including osteocytes, of a 37kDa protein that reacts with the monoclonal anti-FXIII-A antibody AC-1A1. This protein was presumed to be a catalytically active fragment of FXIII-A-83 and to play a major role in bone deposition. We detected a 37kDa AC-1A1 reactive protein in FXIII-A mRNA negative cell lines and in tissues from FXIII-A-/- mice. By mass spectrometric sequencing of AC-1A1 immunoprecipitates, we identified this protein as transaldolase-1, and confirmed that recombinant transaldolase-1 is recognised by AC-1A1. We have also shown that bone deposition is normal in FXIII-A-/-.TG2-/- double knockout mice, casting doubt on the role of transglutaminases in bone mineralisation. Various studies have used antibody AC-1A1 for immunohistochemistry or immunofluorescence. We observe strong FXIII-A dependent staining in paraffin embedded mouse heart sections, with relatively low background in non-expressing mouse cells. In contrast, FXIII-A independent staining predominates in cultured human cells using a standard immunofluorescence procedure. Immunofluorescence is present in membrane compartments that are expected to lack transaldolase, indicating that other off-target antigens are recognised by AC-1A1. This has significant implications for studies that have used this approach to define the subcellular trafficking of FXIII-A in osteocytes.

AB - Transglutaminase activity has been widely implicated in bone deposition. A predominant role has been proposed for factor (F)XIII-A and a subsidiary role suggested for the homologous protein, transglutaminase 2. Full-length FXIII-A is an 83kDa protransglutaminase that is present both in plasma and also in haematopoietic and connective tissue lineages. Several studies have reported expression in murine cells, including osteocytes, of a 37kDa protein that reacts with the monoclonal anti-FXIII-A antibody AC-1A1. This protein was presumed to be a catalytically active fragment of FXIII-A-83 and to play a major role in bone deposition. We detected a 37kDa AC-1A1 reactive protein in FXIII-A mRNA negative cell lines and in tissues from FXIII-A-/- mice. By mass spectrometric sequencing of AC-1A1 immunoprecipitates, we identified this protein as transaldolase-1, and confirmed that recombinant transaldolase-1 is recognised by AC-1A1. We have also shown that bone deposition is normal in FXIII-A-/-.TG2-/- double knockout mice, casting doubt on the role of transglutaminases in bone mineralisation. Various studies have used antibody AC-1A1 for immunohistochemistry or immunofluorescence. We observe strong FXIII-A dependent staining in paraffin embedded mouse heart sections, with relatively low background in non-expressing mouse cells. In contrast, FXIII-A independent staining predominates in cultured human cells using a standard immunofluorescence procedure. Immunofluorescence is present in membrane compartments that are expected to lack transaldolase, indicating that other off-target antigens are recognised by AC-1A1. This has significant implications for studies that have used this approach to define the subcellular trafficking of FXIII-A in osteocytes.

KW - Antibodies

KW - Bone development

KW - FXIII-A

KW - Mice

KW - Transaldolase

KW - Transglutaminases

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U2 - 10.1016/j.matbio.2015.02.001

DO - 10.1016/j.matbio.2015.02.001

M3 - Article

C2 - 25680676

AN - SCOPUS:84930085395

SN - 0945-053X

VL - 43

SP - 85

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JO - Matrix Biology

JF - Matrix Biology

ER -

Normal bone deposition occurs in mice deficient in Factor XIII-A and Transglutaminase 2

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Keywords

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