Identification of a unique form of protein C in the ovine fetus: Developmentally linked transition to the adult form

Marilyn J. Manco-Johnson, Stephen Spedale, Marjolein Peters, Susan F. Townsend, Linda J. Jacobson, Julie Christian, Scott D. Krugman, William W. Hay, John W. Sparks

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

To investigate fetal development of protein C, a pregnant ovine model was used. Protein C was isolated from ovine plasma, and a polyclonal antibody was raised. Citrated plasma was obtained from undisturbed chronically catheterized fetal lambs. On Western blot, nonreduced adult ovine protein C had a molecular mass of 70 kD. Fetal ovine protein C was determined to have a molecular mass of 4 to 6 kD larger than the adult molecule. Crossed Immunoelectrophoresis demonstrated slightly increased anodal migration of the fetal form. Isoelectric focusing demonstrated a decreased pi of the fetal molecule (4.45 versus 4.6). The ovine protein C molecules were deglycosylated with N-glycanase. Deglycosylated fetal protein C migrated more similarly to the adult form, although a portion of the fetal form persisted. These experiments demonstrate the first example of a unique fetal form of a vitamin K-dependent protein and are compatible with increased glycosylation of fetal ovine protein C. It is speculated that altered posttranslational processing may exist as a general process by which certain coagulation proteins are modified during fetal development. mRNA was isolated from maternal and fetal hepatic tissue and analyzed by Northern hybridization. Fetal plasma concentration and hepatic mRNA for protein C were both 40% of normal maternal values from midgestation onward. At term, protein C mRNA increased to adult range (p < 0.025), although plasma protein C concentration decreased slightly (p < 0.001). A transition from fetal to adult protein C form was found beginning 6 d before term birth, with a doubling time of 24 h. These data are compatible with a gestationally determined maturation of ovine protein C. There was no evidence for very low plasma concentrations of protein C during normal fetal and neonatal development. Decreased plasma protein C concentration after birth associated with increased hepatic mRNA suggests increased turnover of protein C in the perinatal period. Further investigation of the transition from fetal to adult forms of protein C and altered hepatic mRNA expression will be necessary to understand, treat, and prevent complications of protein C deficiency in the neonate.

Original languageEnglish (US)
Pages (from-to)365-372
Number of pages8
JournalPediatric Research
Volume37
Issue number3
DOIs
StatePublished - Mar 1995

Fingerprint

Protein C
Sheep
Fetus
Fetal Proteins
Messenger RNA
Fetal Development
Blood Proteins
Liver
Mothers
Protein C Deficiency
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Term Birth
Two-Dimensional Immunoelectrophoresis
Vitamin K
Isoelectric Focusing
Glycosylation
Reference Values

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health

Cite this

Identification of a unique form of protein C in the ovine fetus : Developmentally linked transition to the adult form. / Manco-Johnson, Marilyn J.; Spedale, Stephen; Peters, Marjolein; Townsend, Susan F.; Jacobson, Linda J.; Christian, Julie; Krugman, Scott D.; Hay, William W.; Sparks, John W.

In: Pediatric Research, Vol. 37, No. 3, 03.1995, p. 365-372.

Research output: Contribution to journalArticle

Manco-Johnson, MJ, Spedale, S, Peters, M, Townsend, SF, Jacobson, LJ, Christian, J, Krugman, SD, Hay, WW & Sparks, JW 1995, 'Identification of a unique form of protein C in the ovine fetus: Developmentally linked transition to the adult form', Pediatric Research, vol. 37, no. 3, pp. 365-372. https://doi.org/10.1203/00006450-199503000-00019
Manco-Johnson, Marilyn J. ; Spedale, Stephen ; Peters, Marjolein ; Townsend, Susan F. ; Jacobson, Linda J. ; Christian, Julie ; Krugman, Scott D. ; Hay, William W. ; Sparks, John W. / Identification of a unique form of protein C in the ovine fetus : Developmentally linked transition to the adult form. In: Pediatric Research. 1995 ; Vol. 37, No. 3. pp. 365-372.
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abstract = "To investigate fetal development of protein C, a pregnant ovine model was used. Protein C was isolated from ovine plasma, and a polyclonal antibody was raised. Citrated plasma was obtained from undisturbed chronically catheterized fetal lambs. On Western blot, nonreduced adult ovine protein C had a molecular mass of 70 kD. Fetal ovine protein C was determined to have a molecular mass of 4 to 6 kD larger than the adult molecule. Crossed Immunoelectrophoresis demonstrated slightly increased anodal migration of the fetal form. Isoelectric focusing demonstrated a decreased pi of the fetal molecule (4.45 versus 4.6). The ovine protein C molecules were deglycosylated with N-glycanase. Deglycosylated fetal protein C migrated more similarly to the adult form, although a portion of the fetal form persisted. These experiments demonstrate the first example of a unique fetal form of a vitamin K-dependent protein and are compatible with increased glycosylation of fetal ovine protein C. It is speculated that altered posttranslational processing may exist as a general process by which certain coagulation proteins are modified during fetal development. mRNA was isolated from maternal and fetal hepatic tissue and analyzed by Northern hybridization. Fetal plasma concentration and hepatic mRNA for protein C were both 40{\%} of normal maternal values from midgestation onward. At term, protein C mRNA increased to adult range (p < 0.025), although plasma protein C concentration decreased slightly (p < 0.001). A transition from fetal to adult protein C form was found beginning 6 d before term birth, with a doubling time of 24 h. These data are compatible with a gestationally determined maturation of ovine protein C. There was no evidence for very low plasma concentrations of protein C during normal fetal and neonatal development. Decreased plasma protein C concentration after birth associated with increased hepatic mRNA suggests increased turnover of protein C in the perinatal period. Further investigation of the transition from fetal to adult forms of protein C and altered hepatic mRNA expression will be necessary to understand, treat, and prevent complications of protein C deficiency in the neonate.",
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