TY - JOUR
T1 - Murine Heparin Cofactor II
T2 - Purification, cDNA Sequence, Expression, and Gene Structure
AU - Zhang, Guang Sen
AU - Mehringer, Julie H.
AU - Van Deerlin, Vivianna M.D.
AU - Tollefsen, Douglas M.
AU - Kozak, Christine A.
PY - 1994/3/1
Y1 - 1994/3/1
N2 - Heparin cofactor II (HCII) is a glycoprotein in human plasma that inhibits thrombin rapidly in the presence of dermatan sulfate or heparin. Unexpectedly, we found that HCII activity in murine plasma is present in two proteins of 68 and 72 kDa. The two proteins have the same N-terminal amino acid sequence, and both react with an antibody raised against the C-terminal nine amino acid residues of murine HCII predicted from the cDNA sequence. Treatment of the two proteins with peptide-N4-(N-acetyl-β-glucosaminyl)asparagine amidase yields a single 54-kDa band. Thus, murine plasma contains two forms of HCII that appear to have identical amino acid sequences but differ in the composition of their N-linked oligosaccharides. HCII cDNA clones isolated from a murine liver library include a 1434 bp open reading frame following the first Met codon, a TAA stop codon, and 580 bp of 3′-untranslated sequence terminating in a poly(A) tail. The amino acid sequence deduced from the cDNA contains the N-terminal sequence of purified murine plasma HCII preceded by a 23-residue hydrophobic sequence presumed to be the signal peptide. The amino acid sequence of murine HCII is 87% identical to that of human HCII, the greatest variability occurring in the N-terminal portion of the protein. Northern blot analysis reveals a 2.3-kb HCII mRNA in murine and human liver, but no HCII mRNA is detectable in heart, brain, spleen, lung, skeletal muscle, kidney, testis, placenta, pancreas, or intestine. Southern blot analysis of restriction fragment length polymorphisms in progeny of interspecific and intersubspecific crosses indicates that mice have a single HCII gene (designated Hcf2), which maps to chromosome 16 between Prm-1 and Igl. The murine HCII gene is ∼7.1 kb in size and consists of at least four exons and three introns. The intron/exon origanization is identical to that of the human HCII gene except at the 5′ end, where the murine gene may lack a large intron in the 5′-untranslated region. Our results indicate that HCII is more highly conserved than the human and murine homologues of other serpins such as α1-antitrypsin and α1-antichymotrypsin.
AB - Heparin cofactor II (HCII) is a glycoprotein in human plasma that inhibits thrombin rapidly in the presence of dermatan sulfate or heparin. Unexpectedly, we found that HCII activity in murine plasma is present in two proteins of 68 and 72 kDa. The two proteins have the same N-terminal amino acid sequence, and both react with an antibody raised against the C-terminal nine amino acid residues of murine HCII predicted from the cDNA sequence. Treatment of the two proteins with peptide-N4-(N-acetyl-β-glucosaminyl)asparagine amidase yields a single 54-kDa band. Thus, murine plasma contains two forms of HCII that appear to have identical amino acid sequences but differ in the composition of their N-linked oligosaccharides. HCII cDNA clones isolated from a murine liver library include a 1434 bp open reading frame following the first Met codon, a TAA stop codon, and 580 bp of 3′-untranslated sequence terminating in a poly(A) tail. The amino acid sequence deduced from the cDNA contains the N-terminal sequence of purified murine plasma HCII preceded by a 23-residue hydrophobic sequence presumed to be the signal peptide. The amino acid sequence of murine HCII is 87% identical to that of human HCII, the greatest variability occurring in the N-terminal portion of the protein. Northern blot analysis reveals a 2.3-kb HCII mRNA in murine and human liver, but no HCII mRNA is detectable in heart, brain, spleen, lung, skeletal muscle, kidney, testis, placenta, pancreas, or intestine. Southern blot analysis of restriction fragment length polymorphisms in progeny of interspecific and intersubspecific crosses indicates that mice have a single HCII gene (designated Hcf2), which maps to chromosome 16 between Prm-1 and Igl. The murine HCII gene is ∼7.1 kb in size and consists of at least four exons and three introns. The intron/exon origanization is identical to that of the human HCII gene except at the 5′ end, where the murine gene may lack a large intron in the 5′-untranslated region. Our results indicate that HCII is more highly conserved than the human and murine homologues of other serpins such as α1-antitrypsin and α1-antichymotrypsin.
UR - http://www.scopus.com/inward/record.url?scp=0028281745&partnerID=8YFLogxK
U2 - 10.1021/bi00178a021
DO - 10.1021/bi00178a021
M3 - Article
C2 - 7908224
AN - SCOPUS:0028281745
SN - 0006-2960
VL - 33
SP - 3632
EP - 3642
JO - Biochemistry
JF - Biochemistry
IS - 12
ER -