Propeptide of human protein C is necessary for. gamma. -carboxylation

Journal Article · Tue Nov 03 04:00:00 EST 1987 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00396a022· OSTI ID:5515588

Foster, D C; Rudinski, M S; Schach, B G; Berkner, K L; Kumar, A A; Hagen, F S; Sprecher, C A; Insley, M Y; Davie, E W

Protein C is one of a family of vitamin K dependent proteins, including blood coagulation factors and bone proteins, that contains ..gamma..-carboxyglutamic acid. Sequence analysis of the cDNAs for these proteins has revealed the presence of a prepro leader sequence that contains a pre sequence or hydrophobic signal sequence and a propeptide containing a number of highly conserved amino acids. The pre region is removed from the growing polypeptide chain by signal peptidase, while the pro region is subsequently removed from the protein prior to secretion. In the present study, deletion mutants have been constructed in the propeptide region of the cDNA for human protein C, and the cDNAs were then expressed in mammalian cell culture. These deletions included the removal of 4, 9, 12, 15, 16, or 17 amino acids comprising the carboxyl end of the leader sequence of 42 amino acids. The mutant proteins were then examined by Western blotting, barium citrate adsorption and precipitation, amino acid sequence analysis, and biological activity and compared with the native protein present in normal plasma. These experiments have shown that protein C is readily synthesized in mammalian cell cultures, processed, and secreted as a two-chain molecule with biological activity. Furthermore, the pre portion or signal sequence in human protein C is 18 amino acids in length, and the pro portion of the leader sequence is 24 amino acids in length. Also, during biosynthesis and secretion, the amino-terminal region of the propeptide (residues from about -12 through -17) is important for ..gamma..-carboxylation of protein C, while the present data and those of others indicate that the carboxyl-terminal portion of propeptide (residues -1 through -4) is important for the removal of the pro leader sequence by proteolytic processing.

• Research Organization: Univ. of Washington, Seattle
• OSTI ID: 5515588
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 26:22; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
AMINO ACIDS
BETA DECAY RADIOISOTOPES
BIOSYNTHESIS
BLOOD COAGULATION FACTORS
CARBOXYLATION
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
COAGULANTS
DAYS LIVING RADIOISOTOPES
DNA
DNA SEQUENCING
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
ENZYME IMMUNOASSAY
ENZYMES
GLUTAMIC ACID
HEMATOLOGIC AGENTS
HYDROLASES
IMMUNOASSAY
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
MOLECULAR STRUCTURE
MUTANTS
NUCLEI
NUCLEIC ACIDS
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
PEPTIDE HYDROLASES
RADIOISOTOPES
RECOMBINANT DNA
SECRETION
SERINE PROTEINASES
STRUCTURAL CHEMICAL ANALYSIS
SYNTHESIS