Occurrence of a multimeric high-molecular-weight glyceraldehyde-3-phosphate dehydrogenase in human serum

Rani Kunjithapatham, Jean Francois Geschwind, Lauren Devine, Tatiana N. Boronina, Robert N. Omeally, Robert N. Cole, Michael Torbenson, Shanmugasundaram Ganapathy-Kanniappan

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Cellular glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a phylogenetically conserved, ubiquitous enzyme that plays an indispensable role in energy metabolism. Although a wealth of information is available on cellular GAPDH, there is a clear paucity of data on its extracellular counterpart (i.e., the secreted or extracellular GAPDH). Here, we show that the extracellular GAPDH in human serum is a multimeric, high-molecular-weight, yet glycolytically active enzyme. The high-molecular-weight multimers of serum GAPDH were identified by immunodetection on one- and two-dimensional gel electrophoresis using multiple antibodies specific for various epitopes of GAPDH. Partial purification of serum GAPDH by DEAE Affigel affinity/ion exchange chromatography further established the multimeric composition of serum GAPDH. In vitro data demonstrated that human cell lines secrete a multimeric, high-molecular-weight enzyme similar to that of serum GAPDH. Furthermore, LC-MS/MS analysis of extracellular GAPDH from human cell lines confirmed the presence of unique peptides of GAPDH in the high-molecular-weight subunits. Furthermore, data from pulse-chase experiments established the presence of high-molecular-weight subunits in the secreted, extracellular GAPDH. Taken together, our findings demonstrate the presence of a high-molecular-weight, enzymatically active secretory GAPDH in human serum that may have a hitherto unknown function in humans.

Original languageEnglish (US)
Pages (from-to)1645-1656
Number of pages12
JournalJournal of Proteome Research
Volume14
Issue number4
DOIs
StatePublished - Apr 3 2015
Externally publishedYes

Fingerprint

Glyceraldehyde-3-Phosphate Dehydrogenases
Molecular Weight
Molecular weight
Serum
Enzymes
Cells
Cell Line
Ion Exchange Chromatography
Electrophoresis, Gel, Two-Dimensional
Chromatography
Electrophoresis
Energy Metabolism
Purification
Epitopes
Ion exchange

Keywords

  • 2D gel electrophoresis
  • DEAE Affigel Blue column chromatography
  • extracellular GAPDH
  • multimer
  • serum GAPDH

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Kunjithapatham, R., Geschwind, J. F., Devine, L., Boronina, T. N., Omeally, R. N., Cole, R. N., ... Ganapathy-Kanniappan, S. (2015). Occurrence of a multimeric high-molecular-weight glyceraldehyde-3-phosphate dehydrogenase in human serum. Journal of Proteome Research, 14(4), 1645-1656. https://doi.org/10.1021/acs.jproteome.5b00089

Occurrence of a multimeric high-molecular-weight glyceraldehyde-3-phosphate dehydrogenase in human serum. / Kunjithapatham, Rani; Geschwind, Jean Francois; Devine, Lauren; Boronina, Tatiana N.; Omeally, Robert N.; Cole, Robert N.; Torbenson, Michael; Ganapathy-Kanniappan, Shanmugasundaram.

In: Journal of Proteome Research, Vol. 14, No. 4, 03.04.2015, p. 1645-1656.

Research output: Contribution to journalArticle

Kunjithapatham, R, Geschwind, JF, Devine, L, Boronina, TN, Omeally, RN, Cole, RN, Torbenson, M & Ganapathy-Kanniappan, S 2015, 'Occurrence of a multimeric high-molecular-weight glyceraldehyde-3-phosphate dehydrogenase in human serum', Journal of Proteome Research, vol. 14, no. 4, pp. 1645-1656. https://doi.org/10.1021/acs.jproteome.5b00089
Kunjithapatham, Rani ; Geschwind, Jean Francois ; Devine, Lauren ; Boronina, Tatiana N. ; Omeally, Robert N. ; Cole, Robert N. ; Torbenson, Michael ; Ganapathy-Kanniappan, Shanmugasundaram. / Occurrence of a multimeric high-molecular-weight glyceraldehyde-3-phosphate dehydrogenase in human serum. In: Journal of Proteome Research. 2015 ; Vol. 14, No. 4. pp. 1645-1656.
@article{fe4e38d893c74c97b32809e747f2938a,
title = "Occurrence of a multimeric high-molecular-weight glyceraldehyde-3-phosphate dehydrogenase in human serum",
abstract = "Cellular glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a phylogenetically conserved, ubiquitous enzyme that plays an indispensable role in energy metabolism. Although a wealth of information is available on cellular GAPDH, there is a clear paucity of data on its extracellular counterpart (i.e., the secreted or extracellular GAPDH). Here, we show that the extracellular GAPDH in human serum is a multimeric, high-molecular-weight, yet glycolytically active enzyme. The high-molecular-weight multimers of serum GAPDH were identified by immunodetection on one- and two-dimensional gel electrophoresis using multiple antibodies specific for various epitopes of GAPDH. Partial purification of serum GAPDH by DEAE Affigel affinity/ion exchange chromatography further established the multimeric composition of serum GAPDH. In vitro data demonstrated that human cell lines secrete a multimeric, high-molecular-weight enzyme similar to that of serum GAPDH. Furthermore, LC-MS/MS analysis of extracellular GAPDH from human cell lines confirmed the presence of unique peptides of GAPDH in the high-molecular-weight subunits. Furthermore, data from pulse-chase experiments established the presence of high-molecular-weight subunits in the secreted, extracellular GAPDH. Taken together, our findings demonstrate the presence of a high-molecular-weight, enzymatically active secretory GAPDH in human serum that may have a hitherto unknown function in humans.",
keywords = "2D gel electrophoresis, DEAE Affigel Blue column chromatography, extracellular GAPDH, multimer, serum GAPDH",
author = "Rani Kunjithapatham and Geschwind, {Jean Francois} and Lauren Devine and Boronina, {Tatiana N.} and Omeally, {Robert N.} and Cole, {Robert N.} and Michael Torbenson and Shanmugasundaram Ganapathy-Kanniappan",
year = "2015",
month = "4",
day = "3",
doi = "10.1021/acs.jproteome.5b00089",
language = "English (US)",
volume = "14",
pages = "1645--1656",
journal = "Journal of Proteome Research",
issn = "1535-3893",
publisher = "American Chemical Society",
number = "4",

}

TY - JOUR

T1 - Occurrence of a multimeric high-molecular-weight glyceraldehyde-3-phosphate dehydrogenase in human serum

AU - Kunjithapatham, Rani

AU - Geschwind, Jean Francois

AU - Devine, Lauren

AU - Boronina, Tatiana N.

AU - Omeally, Robert N.

AU - Cole, Robert N.

AU - Torbenson, Michael

AU - Ganapathy-Kanniappan, Shanmugasundaram

PY - 2015/4/3

Y1 - 2015/4/3

N2 - Cellular glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a phylogenetically conserved, ubiquitous enzyme that plays an indispensable role in energy metabolism. Although a wealth of information is available on cellular GAPDH, there is a clear paucity of data on its extracellular counterpart (i.e., the secreted or extracellular GAPDH). Here, we show that the extracellular GAPDH in human serum is a multimeric, high-molecular-weight, yet glycolytically active enzyme. The high-molecular-weight multimers of serum GAPDH were identified by immunodetection on one- and two-dimensional gel electrophoresis using multiple antibodies specific for various epitopes of GAPDH. Partial purification of serum GAPDH by DEAE Affigel affinity/ion exchange chromatography further established the multimeric composition of serum GAPDH. In vitro data demonstrated that human cell lines secrete a multimeric, high-molecular-weight enzyme similar to that of serum GAPDH. Furthermore, LC-MS/MS analysis of extracellular GAPDH from human cell lines confirmed the presence of unique peptides of GAPDH in the high-molecular-weight subunits. Furthermore, data from pulse-chase experiments established the presence of high-molecular-weight subunits in the secreted, extracellular GAPDH. Taken together, our findings demonstrate the presence of a high-molecular-weight, enzymatically active secretory GAPDH in human serum that may have a hitherto unknown function in humans.

AB - Cellular glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a phylogenetically conserved, ubiquitous enzyme that plays an indispensable role in energy metabolism. Although a wealth of information is available on cellular GAPDH, there is a clear paucity of data on its extracellular counterpart (i.e., the secreted or extracellular GAPDH). Here, we show that the extracellular GAPDH in human serum is a multimeric, high-molecular-weight, yet glycolytically active enzyme. The high-molecular-weight multimers of serum GAPDH were identified by immunodetection on one- and two-dimensional gel electrophoresis using multiple antibodies specific for various epitopes of GAPDH. Partial purification of serum GAPDH by DEAE Affigel affinity/ion exchange chromatography further established the multimeric composition of serum GAPDH. In vitro data demonstrated that human cell lines secrete a multimeric, high-molecular-weight enzyme similar to that of serum GAPDH. Furthermore, LC-MS/MS analysis of extracellular GAPDH from human cell lines confirmed the presence of unique peptides of GAPDH in the high-molecular-weight subunits. Furthermore, data from pulse-chase experiments established the presence of high-molecular-weight subunits in the secreted, extracellular GAPDH. Taken together, our findings demonstrate the presence of a high-molecular-weight, enzymatically active secretory GAPDH in human serum that may have a hitherto unknown function in humans.

KW - 2D gel electrophoresis

KW - DEAE Affigel Blue column chromatography

KW - extracellular GAPDH

KW - multimer

KW - serum GAPDH

UR - http://www.scopus.com/inward/record.url?scp=84926442812&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84926442812&partnerID=8YFLogxK

U2 - 10.1021/acs.jproteome.5b00089

DO - 10.1021/acs.jproteome.5b00089

M3 - Article

C2 - 25734908

AN - SCOPUS:84926442812

VL - 14

SP - 1645

EP - 1656

JO - Journal of Proteome Research

JF - Journal of Proteome Research

SN - 1535-3893

IS - 4

ER -