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Thromb Haemost 2001; 86(04): 1017-1022
DOI: 10.1055/s-0037-1616527
Special Article
Schattauer GmbH

Abnormal Propeptide Processing Resulting in the Presence of Two Abnormal Species of Protein C in Plasma

Characterization of the Dysfunctional Protein C Padua3 (Protein CR-1L/propeptide)
Paolo Simioni
1   Department of Medical and Surgical Sciences, Second Chair of Medicine, University of Padua, Padua Italy
,
Michael Kalafatis
2   Department of Chemistry, Cleveland State University, Cleveland, OH, USA
3   Department of Molecular Cardiology, Cleveland Clinic Foundation, Cleveland, OH, USA
,
Daniela Tormene
1   Department of Medical and Surgical Sciences, Second Chair of Medicine, University of Padua, Padua Italy
,
Sonia Luni
1   Department of Medical and Surgical Sciences, Second Chair of Medicine, University of Padua, Padua Italy
,
Patrizia Zerbinati
1   Department of Medical and Surgical Sciences, Second Chair of Medicine, University of Padua, Padua Italy
,
Luisa Barzon
4   Department of Microbiology, University of Padua, Padua, Italy
,
Giorgio Palù
4   Department of Microbiology, University of Padua, Padua, Italy
,
Antonio Girolami
1   Department of Medical and Surgical Sciences, Second Chair of Medicine, University of Padua, Padua Italy
› Author AffiliationsSupported by funds from the Department of Medical and Surgical Sciences, University of Padua (to P.S.) and by Start-up-funds from the Chemistry Department at Cleveland State University (to M.K.).
Further Information

Publication History

Received 08 January 2001

Accepted after resubmission 16 May 2001

Publication Date:
09 December 2017 (online)

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Summary

A heterozygous GT transversion at position 1388 of the protein C (PC) gene which predicted the substitution of Arg-1 to a Leu (PCR-1L) was identified in a thrombophilic patient. The PCR-1L was purified from the patient’s plasma by immunoaffinity chromatography using Ca++-independent and Ca++-dependent monoclonal antibodies. NH2-terminal sequencing of the light chain of PCR-1L revealed two amino acid sequences: one was identical to the complete propeptide sequence of PC, while the other matched the normal PC light chain sequence elongated by one amino acid (Leucine at position 1). Activated PCR-1L/propeptide exhibited normal amidolytic and impaired anticoagulant activity. Thus, the substitution of a Leu for an Arg at position -1 of PC shifts the propeptidase cleavage site by one amino acid. In addition, in PCR-1L/propeptide the propeptide cleavage at Lys-2 is less efficient since approximately 60% of PC variant molecules present in patient’s plasma retained the entire propeptide. Our findings suggest that depending on the specific amino acid substitution at position-1, PC can be secreted in plasma containing the entire propeptide attached to the light chain. Impaired interaction of elongated APC molecules with a membrane-surface and/or factor Va which is the physiological substrate for APC, is manifested in vivo by thrombophilia.

Keywords

Dysfunctional protein C - type II defect - propeptide clearage - thrombophilia - factor V
 

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