ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets

Paul W. Wright, Laura C. Bolling, Meredith E. Calvert, Olga F. Sarmento, Elizabeth V. Berkeley, Margaret C. Shea, Zhonglin Hao, Friederike C. Jayes, Leigh Ann Bush, Jagathpala Shetty, Amy N. Shore, Prabhakara P. Reddi, Kenneth S. Tung, Eileen Samy, Margaretta M. Allietta, Nicholas E. Sherman, John C. Herr, Scott A. Coonrod

Research output: Contribution to journalArticle

100 Citations (Scopus)

Abstract

Selected for its high relative abundance, a protein spot of MW ∼75 kDa, pI 5.5 was cored from a Coomassie-stained two-dimensional gel of proteins from 2850 zona-free metaphase II mouse eggs and analyzed by tandem mass spectrometry (TMS), and novel microsequences were identified that indicated a previously uncharacterized egg protein. A 2.4-kb cDNA was then amplified from a mouse ovarian adapter-ligated cDNA library by RACE-PCR, and a unique 2043-bp open reading frame was defined encoding a 681-amino-acid protein. Comparison of the deduced amino acid sequence with the nonredundant database demonstrated that the protein was ∼40% identical to the calcium-dependent peptidylarginine deiminase (PAD) enzyme family. Northern blotting, RT-PCR, and in situ hybridization analyses indicated that the protein was abundantly expressed in the ovary, weakly expressed in the testis, and absent from other tissues. Based on the homology with PADs and its oocyte-abundant expression pattern, the protein was designated ePAD, for egg and embryo-abundant peptidylarginine deiminase-like protein. Anti-recombinant ePAD monospecific antibodies localized the molecule to the cytoplasm of oocytes in primordial, primary, secondary, and Graafian follicles in ovarian sections, while no other ovarian cell type was stained. ePAD was also expressed in the immature oocyte, mature egg, and through the blastocyst stage of embryonic development, where expression levels began to decrease. Immunoelectron microscopy localized ePAD to egg cytoplasmic sheets, a unique keratin-containing intermediate filament structure found only in mammalian eggs and in early embryos, and known to undergo reorganization at critical stages of development. Previous reports that PAD-mediated deimination of epithelial cell keratin results in cytoskeletal remodeling suggest a possible role for ePAD in cytoskeletal reorganization in the egg and early embryo.

Original languageEnglish (US)
Pages (from-to)74-89
Number of pages16
JournalDevelopmental Biology
Volume256
Issue number1
DOIs
StatePublished - Apr 1 2003

Fingerprint

Oocytes
Ovum
Embryonic Structures
Proteins
Keratins
Eggs
Egg Proteins
Polymerase Chain Reaction
Protein Databases
Ovarian Follicle
Intermediate Filaments
Immunoelectron Microscopy
Herpes Zoster
Blastocyst
Metaphase
Tandem Mass Spectrometry
Gene Library
Northern Blotting
Open Reading Frames
Embryonic Development

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

Cite this

Wright, P. W., Bolling, L. C., Calvert, M. E., Sarmento, O. F., Berkeley, E. V., Shea, M. C., ... Coonrod, S. A. (2003). ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets. Developmental Biology, 256(1), 74-89. https://doi.org/10.1016/S0012-1606(02)00126-4

ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets. / Wright, Paul W.; Bolling, Laura C.; Calvert, Meredith E.; Sarmento, Olga F.; Berkeley, Elizabeth V.; Shea, Margaret C.; Hao, Zhonglin; Jayes, Friederike C.; Bush, Leigh Ann; Shetty, Jagathpala; Shore, Amy N.; Reddi, Prabhakara P.; Tung, Kenneth S.; Samy, Eileen; Allietta, Margaretta M.; Sherman, Nicholas E.; Herr, John C.; Coonrod, Scott A.

In: Developmental Biology, Vol. 256, No. 1, 01.04.2003, p. 74-89.

Research output: Contribution to journalArticle

Wright, PW, Bolling, LC, Calvert, ME, Sarmento, OF, Berkeley, EV, Shea, MC, Hao, Z, Jayes, FC, Bush, LA, Shetty, J, Shore, AN, Reddi, PP, Tung, KS, Samy, E, Allietta, MM, Sherman, NE, Herr, JC & Coonrod, SA 2003, 'ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets', Developmental Biology, vol. 256, no. 1, pp. 74-89. https://doi.org/10.1016/S0012-1606(02)00126-4
Wright, Paul W. ; Bolling, Laura C. ; Calvert, Meredith E. ; Sarmento, Olga F. ; Berkeley, Elizabeth V. ; Shea, Margaret C. ; Hao, Zhonglin ; Jayes, Friederike C. ; Bush, Leigh Ann ; Shetty, Jagathpala ; Shore, Amy N. ; Reddi, Prabhakara P. ; Tung, Kenneth S. ; Samy, Eileen ; Allietta, Margaretta M. ; Sherman, Nicholas E. ; Herr, John C. ; Coonrod, Scott A. / ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets. In: Developmental Biology. 2003 ; Vol. 256, No. 1. pp. 74-89.
@article{bf99cd7e76c947548d3f9d0c2f0358f5,
title = "ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets",
abstract = "Selected for its high relative abundance, a protein spot of MW ∼75 kDa, pI 5.5 was cored from a Coomassie-stained two-dimensional gel of proteins from 2850 zona-free metaphase II mouse eggs and analyzed by tandem mass spectrometry (TMS), and novel microsequences were identified that indicated a previously uncharacterized egg protein. A 2.4-kb cDNA was then amplified from a mouse ovarian adapter-ligated cDNA library by RACE-PCR, and a unique 2043-bp open reading frame was defined encoding a 681-amino-acid protein. Comparison of the deduced amino acid sequence with the nonredundant database demonstrated that the protein was ∼40{\%} identical to the calcium-dependent peptidylarginine deiminase (PAD) enzyme family. Northern blotting, RT-PCR, and in situ hybridization analyses indicated that the protein was abundantly expressed in the ovary, weakly expressed in the testis, and absent from other tissues. Based on the homology with PADs and its oocyte-abundant expression pattern, the protein was designated ePAD, for egg and embryo-abundant peptidylarginine deiminase-like protein. Anti-recombinant ePAD monospecific antibodies localized the molecule to the cytoplasm of oocytes in primordial, primary, secondary, and Graafian follicles in ovarian sections, while no other ovarian cell type was stained. ePAD was also expressed in the immature oocyte, mature egg, and through the blastocyst stage of embryonic development, where expression levels began to decrease. Immunoelectron microscopy localized ePAD to egg cytoplasmic sheets, a unique keratin-containing intermediate filament structure found only in mammalian eggs and in early embryos, and known to undergo reorganization at critical stages of development. Previous reports that PAD-mediated deimination of epithelial cell keratin results in cytoskeletal remodeling suggest a possible role for ePAD in cytoskeletal reorganization in the egg and early embryo.",
author = "Wright, {Paul W.} and Bolling, {Laura C.} and Calvert, {Meredith E.} and Sarmento, {Olga F.} and Berkeley, {Elizabeth V.} and Shea, {Margaret C.} and Zhonglin Hao and Jayes, {Friederike C.} and Bush, {Leigh Ann} and Jagathpala Shetty and Shore, {Amy N.} and Reddi, {Prabhakara P.} and Tung, {Kenneth S.} and Eileen Samy and Allietta, {Margaretta M.} and Sherman, {Nicholas E.} and Herr, {John C.} and Coonrod, {Scott A.}",
year = "2003",
month = "4",
day = "1",
doi = "10.1016/S0012-1606(02)00126-4",
language = "English (US)",
volume = "256",
pages = "74--89",
journal = "Developmental Biology",
issn = "0012-1606",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets

AU - Wright, Paul W.

AU - Bolling, Laura C.

AU - Calvert, Meredith E.

AU - Sarmento, Olga F.

AU - Berkeley, Elizabeth V.

AU - Shea, Margaret C.

AU - Hao, Zhonglin

AU - Jayes, Friederike C.

AU - Bush, Leigh Ann

AU - Shetty, Jagathpala

AU - Shore, Amy N.

AU - Reddi, Prabhakara P.

AU - Tung, Kenneth S.

AU - Samy, Eileen

AU - Allietta, Margaretta M.

AU - Sherman, Nicholas E.

AU - Herr, John C.

AU - Coonrod, Scott A.

PY - 2003/4/1

Y1 - 2003/4/1

N2 - Selected for its high relative abundance, a protein spot of MW ∼75 kDa, pI 5.5 was cored from a Coomassie-stained two-dimensional gel of proteins from 2850 zona-free metaphase II mouse eggs and analyzed by tandem mass spectrometry (TMS), and novel microsequences were identified that indicated a previously uncharacterized egg protein. A 2.4-kb cDNA was then amplified from a mouse ovarian adapter-ligated cDNA library by RACE-PCR, and a unique 2043-bp open reading frame was defined encoding a 681-amino-acid protein. Comparison of the deduced amino acid sequence with the nonredundant database demonstrated that the protein was ∼40% identical to the calcium-dependent peptidylarginine deiminase (PAD) enzyme family. Northern blotting, RT-PCR, and in situ hybridization analyses indicated that the protein was abundantly expressed in the ovary, weakly expressed in the testis, and absent from other tissues. Based on the homology with PADs and its oocyte-abundant expression pattern, the protein was designated ePAD, for egg and embryo-abundant peptidylarginine deiminase-like protein. Anti-recombinant ePAD monospecific antibodies localized the molecule to the cytoplasm of oocytes in primordial, primary, secondary, and Graafian follicles in ovarian sections, while no other ovarian cell type was stained. ePAD was also expressed in the immature oocyte, mature egg, and through the blastocyst stage of embryonic development, where expression levels began to decrease. Immunoelectron microscopy localized ePAD to egg cytoplasmic sheets, a unique keratin-containing intermediate filament structure found only in mammalian eggs and in early embryos, and known to undergo reorganization at critical stages of development. Previous reports that PAD-mediated deimination of epithelial cell keratin results in cytoskeletal remodeling suggest a possible role for ePAD in cytoskeletal reorganization in the egg and early embryo.

AB - Selected for its high relative abundance, a protein spot of MW ∼75 kDa, pI 5.5 was cored from a Coomassie-stained two-dimensional gel of proteins from 2850 zona-free metaphase II mouse eggs and analyzed by tandem mass spectrometry (TMS), and novel microsequences were identified that indicated a previously uncharacterized egg protein. A 2.4-kb cDNA was then amplified from a mouse ovarian adapter-ligated cDNA library by RACE-PCR, and a unique 2043-bp open reading frame was defined encoding a 681-amino-acid protein. Comparison of the deduced amino acid sequence with the nonredundant database demonstrated that the protein was ∼40% identical to the calcium-dependent peptidylarginine deiminase (PAD) enzyme family. Northern blotting, RT-PCR, and in situ hybridization analyses indicated that the protein was abundantly expressed in the ovary, weakly expressed in the testis, and absent from other tissues. Based on the homology with PADs and its oocyte-abundant expression pattern, the protein was designated ePAD, for egg and embryo-abundant peptidylarginine deiminase-like protein. Anti-recombinant ePAD monospecific antibodies localized the molecule to the cytoplasm of oocytes in primordial, primary, secondary, and Graafian follicles in ovarian sections, while no other ovarian cell type was stained. ePAD was also expressed in the immature oocyte, mature egg, and through the blastocyst stage of embryonic development, where expression levels began to decrease. Immunoelectron microscopy localized ePAD to egg cytoplasmic sheets, a unique keratin-containing intermediate filament structure found only in mammalian eggs and in early embryos, and known to undergo reorganization at critical stages of development. Previous reports that PAD-mediated deimination of epithelial cell keratin results in cytoskeletal remodeling suggest a possible role for ePAD in cytoskeletal reorganization in the egg and early embryo.

UR - http://www.scopus.com/inward/record.url?scp=0037376592&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037376592&partnerID=8YFLogxK

U2 - 10.1016/S0012-1606(02)00126-4

DO - 10.1016/S0012-1606(02)00126-4

M3 - Article

VL - 256

SP - 74

EP - 89

JO - Developmental Biology

JF - Developmental Biology

SN - 0012-1606

IS - 1

ER -