scholarly journals Characterization of primary amino acid sequence of human complement control protein factor I from an analysis of cDNA clones

1987 ◽  
Vol 242 (3) ◽  
pp. 849-856 ◽  
Author(s):  
C F Catterall ◽  
A Lyons ◽  
R B Sim ◽  
A J Day ◽  
T J R Harris
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Heavy Chain ◽  
Light Chain ◽  
Cdna Clones ◽  
Serine Proteinases ◽  
Human Complement ◽  
Factor I ◽  
Protein Factor ◽  
Control Protein

A cDNA clone of the mRNA coding for the human complement system control protein Factor I has been isolated. The predicted amino acid sequence obtained from the DNA sequence demonstrates a protein consisting of a heavy chain (Mr 35,400) linked to a light chain (Mr 27,600), both of which contain three sites for N-linked glycosylation. The light chain has clear homology with other serine proteinases, most notably in the region of the catalytically active and structurally important amino acids and shares some of the features characteristic of the plasminogen activators. The heavy chain has a clear ‘mosaic’ nature typical of the plasma serine proteinases; in particular it contains class A and class B LDL (low-density lipoprotein) receptor repeats with conserved cysteine residues similar to those found in other complement proteins.

scholarly journals Arrangement of the disulphide bridges in human low-Mr kininogen

1987 ◽  
Vol 247 (1) ◽  
pp. 15-21 ◽  
Author(s):  
J Kellermann ◽  
C Thelen ◽  
F Lottspeich ◽  
A Henschen ◽  
R Vogel ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Heavy Chain ◽  
Light Chain ◽  
The Other ◽  
Critical Importance

The arrangement of the disulphide bridges in human low-Mr kininogen has been elucidated. Low-Mr kininogen contains 18 half-cystine residues forming nine disulphide bridges. The first and the last half-cystine residues of the amino acid sequence form a disulphide loop which spans the heavy- and the light-chain portion of the kininogen molecule. The other 16 half-cystine residues are linked consecutively to form eight loops of 4-20 amino acids; these loops are lined up in the heavy-chain portion of the kininogen molecule. In this way, a particular pattern of disulphide loops is formed which seems to be of critical importance for the inhibitor function of human kininogen.

scholarly journals Cellular myosin heavy chain in human leukocytes: isolation of 5' cDNA clones, characterization of the protein, chromosomal localization, and upregulation during myeloid differentiation

Blood ◽  
1991 ◽  
Vol 78 (7) ◽  
pp. 1826-1833 ◽  
Author(s):  
LE Toothaker ◽  
DA Gonzalez ◽  
N Tung ◽  
RS Lemons ◽  
MM Le Beau ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Myosin Heavy Chain ◽  
Cell Lines ◽  
Heavy Chain ◽  
Myeloid Cell ◽  
Predicted Amino Acid Sequence ◽  
Cdna Clones ◽  
Granulocytic Differentiation ◽  
Human Leukocytes

Abstract We have isolated 5′ cDNA clones encoding a member of the cellular myosin heavy chain gene family from human leukocytes. The predicted amino acid sequence shows 93% identity to a chicken cellular myosin heavy chain, 76% to chicken smooth muscle, and 40% to human sarcomeric myosin heavy chain. The mRNA is expressed as a 7.4- to 7.9-kb doublet in many nonmuscle cells, and is upregulated in myeloid cell lines on induction from a proliferating to a differentiated state. Antisera raised against a peptide made from the predicted amino acid sequence specifically reacts with a 224-Kd polypeptide in leukocyte cell lines, and the protein is also upregulated during the induction of monocytic and granulocytic differentiation in these cells. The gene for this cellular myosin heavy chain maps to chromosome 22, bands q12.3-q13.1, demonstrating that it is not located in the previously described sarcomeric gene clusters on chromosomes 14 and 17. This cellular myosin heavy chain may be a major contractile protein responsible for movement in myeloid cell lines because no mRNA for sarcomeric myosin heavy chain is detected in these cells.

Cl̄r and Cl̄s subcomponents of human complement: Two serine proteinases lacking the ‘histidine-loop’ disulphide bridge

1981 ◽  
Vol 1 (10) ◽  
pp. 779-784 ◽  
Author(s):  
Gerard J. Arlaud ◽  
Jean Gagnon
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Histidine Residue ◽  
Serine Proteinases ◽  
Human Complement ◽  
Disulphide Bridge ◽  
Terminal Amino Acid ◽  
Relay System ◽  
Terminal Amino ◽  
Terminal Amino Acid Sequence

The N-terminal amino acid sequence of human Cl̄s b chain has been extended to 52 residues. The histidine residue involved in the charge-relay system is located at position 38, whereas the ‘histidine-loop’ disulphide bridge is missing. So far, human complement subcomponents Cl̄s are the only known mammalian serine proteinases lacking this disulphide bridge.

scholarly journals Cellular myosin heavy chain in human leukocytes: isolation of 5' cDNA clones, characterization of the protein, chromosomal localization, and upregulation during myeloid differentiation

Blood ◽  
1991 ◽  
Vol 78 (7) ◽  
pp. 1826-1833
Author(s):  
LE Toothaker ◽  
DA Gonzalez ◽  
N Tung ◽  
RS Lemons ◽  
MM Le Beau ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Myosin Heavy Chain ◽  
Cell Lines ◽  
Heavy Chain ◽  
Myeloid Cell ◽  
Predicted Amino Acid Sequence ◽  
Cdna Clones ◽  
Granulocytic Differentiation ◽  
Human Leukocytes

We have isolated 5′ cDNA clones encoding a member of the cellular myosin heavy chain gene family from human leukocytes. The predicted amino acid sequence shows 93% identity to a chicken cellular myosin heavy chain, 76% to chicken smooth muscle, and 40% to human sarcomeric myosin heavy chain. The mRNA is expressed as a 7.4- to 7.9-kb doublet in many nonmuscle cells, and is upregulated in myeloid cell lines on induction from a proliferating to a differentiated state. Antisera raised against a peptide made from the predicted amino acid sequence specifically reacts with a 224-Kd polypeptide in leukocyte cell lines, and the protein is also upregulated during the induction of monocytic and granulocytic differentiation in these cells. The gene for this cellular myosin heavy chain maps to chromosome 22, bands q12.3-q13.1, demonstrating that it is not located in the previously described sarcomeric gene clusters on chromosomes 14 and 17. This cellular myosin heavy chain may be a major contractile protein responsible for movement in myeloid cell lines because no mRNA for sarcomeric myosin heavy chain is detected in these cells.

Amino Acid Sequence of Trocarin, a Prothrombin Activator FromTropidechis carinatus Venom: Its Structural Similarity to Coagulation Factor Xa

Blood ◽  
1999 ◽  
Vol 94 (2) ◽  
pp. 621-631 ◽  
Author(s):  
Jeremiah S. Joseph ◽  
Maxey C.M. Chung ◽  
Kandiah Jeyaseelan ◽  
R. Manjunatha Kini
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Blood Coagulation ◽  
Snake Venom ◽  
Heavy Chain ◽  
Light Chain ◽  
Critical Role ◽  
Factor Xa ◽  
Coagulation Factor ◽  
Group Ii

Among snake venom procoagulant proteins, group II prothrombin activators are functionally similar to blood coagulation factor Xa. We have purified and partially characterized the enzymatic properties of trocarin, the group II prothrombin activator from the venom of the Australian elapid, Tropidechis carinatus (rough-scaled snake). Prothrombin activation by trocarin is enhanced by Ca2+, phospholipids, and factor Va, similar to that by factor Xa. However, its amidolytic activity on peptide substrate S-2222 is significantly lower. We have determined the complete amino acid sequence of trocarin. It is a 46,515-Dalton glycoprotein highly homologous to factor Xa and shares the same domain architecture. The light chain possesses an N-terminal Gla domain containing 11 γ-carboxyglutamic acid residues, followed by two epidermal growth factor (EGF)-like domains; the heavy chain is a serine proteinase. Both chains are likely glycosylated: the light chain at Ser 52 and the heavy chain at Asn 45. Unlike other types of venom procoagulants, trocarin is the first true structural homologue of a coagulation factor. It clots snake plasma and thus may be similar, if not identical, to snake blood coagulation factor Xa. Unlike blood factor Xa, it is expressed in high quantities and in a nonhepatic tissue, making snake venom the richest source of factor Xa-like proteins. It induces cyanosis and death in mice at 1 mg/kg body weight. Thus, trocarin acts as a toxin in venom and a similar, if not identical, protein plays a critical role in hemostasis.

scholarly journals Dictyostelium discoideum myosin: isolation and characterization of cDNAs encoding the essential light chain.

1988 ◽  
Vol 8 (2) ◽  
pp. 794-801 ◽  
Author(s):  
R L Chisholm ◽  
A M Rushforth ◽  
R S Pollenz ◽  
E R Kuczmarski ◽  
S R Tafuri
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Dictyostelium Discoideum ◽  
Light Chain ◽  
Single Gene ◽  
Cdna Expression Library ◽  
Cdna Clones ◽  
Expression Library ◽  
Essential Light Chain ◽  
Isolation And Characterization

We used an antibody specific for Dictyostelium discoideum myosin to screen a lambda gt11 cDNA expression library to obtain cDNA clones which encode the Dictyostelium essential myosin light chain (EMLC). The amino acid sequence predicted from the sequence of the cDNA clone showed 31.5% identity with the amino acid sequence of the chicken EMLC. Comparisons of the Dictyostelium EMLC, a nonmuscle cell type, with EMLC sequences from similar MLCs of skeletal- and smooth-muscle origin, showed distinct regions of homology. Much of the observed homology was localized to regions corresponding to consensus Ca2+-binding of E-F hand domains. Southern blot analysis suggested that the Dictyostelium genome contains a single gene encoding the EMLC. Examination of the pattern of EMLC mRNA expression showed that a significant increase in EMLC message levels occurred during the first few hours of development, coinciding with increased actin expression and immediately preceding the period of maximal chemotactic activity.

MOLECULAR CLONING OF HUMAN TISSUE FACTOR cDNA

1987 ◽  
Author(s):  
T S Edgington ◽  
J H Morrissey ◽  
H Fakhrai
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Tissue Factor ◽  
Heavy Chain ◽  
Human Tissue ◽  
Cell Surface Receptor ◽  
Intact Protein ◽  
Cdna Clones ◽  
Functional Domain ◽  
Amino Terminal

Tissue factor (TF), the cell-surface receptor and allo-steric activator for factor Vll/VIIa, is important in hemostasis andinflammation. The TF apoprotein was purifiedfrom human brain using factor Vll-affinitychromatography and SDS gel electrophoresis,and was found to consist of a 47 kDa heavy chain plus a 12.5 kDa light chain. Approximately one-third of the heavy chain amino acid sequence was determined for four regions by microsequencing the intact protein and peptides derived from V8-protease digestion. A λgtll cDNA library, made from mRNA derived from the human fibroblastic cell line WI38,was screened with (a) affinity-purified rabbit antibodies to human tissue factor, and (b) a 45-mer oligonucleotide probe based on TF heavy chain amino acid sequence. Five overlapping cDNA clones were identifiedand sequenced which confirmed all four partial TF amino acid sequences. Together these clones span the entire heavy chain coding sequence as well as 5" and 3" nontranslated regions. The N-terminusof the TF heavy chain is preceded by an unusually long signal peptide which appears to be cleaved at alternative sites two amino acids apart. This results in two variants of TF heavy chains which differ slightly in length and amino-terminal sequence.The deduced protein sequence shows no major homology to known protein sequences. However,a relatively uncommon tripeptide sequence, Trp-Lys-Ser (WKS), appears three times in the TF heavy chain. This tripeptidesequence also occurs in HMW kininogen, factor VIII,von Willebrand"s factor andant ithrombin-III. Limited sequence similarity is observed in flanking sequences,andthis may indicate a possible functional domain for the recognition of members ofthe vitamin K-dependent serine protease famil.Supported by NIH grants HL-16411 andCA-41085.

Dictyostelium discoideum myosin: isolation and characterization of cDNAs encoding the essential light chain

1988 ◽  
Vol 8 (2) ◽  
pp. 794-801
Author(s):  
R L Chisholm ◽  
A M Rushforth ◽  
R S Pollenz ◽  
E R Kuczmarski ◽  
S R Tafuri
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Dictyostelium Discoideum ◽  
Light Chain ◽  
Single Gene ◽  
Cdna Expression Library ◽  
Cdna Clones ◽  
Expression Library ◽  
Essential Light Chain ◽  
Isolation And Characterization

We used an antibody specific for Dictyostelium discoideum myosin to screen a lambda gt11 cDNA expression library to obtain cDNA clones which encode the Dictyostelium essential myosin light chain (EMLC). The amino acid sequence predicted from the sequence of the cDNA clone showed 31.5% identity with the amino acid sequence of the chicken EMLC. Comparisons of the Dictyostelium EMLC, a nonmuscle cell type, with EMLC sequences from similar MLCs of skeletal- and smooth-muscle origin, showed distinct regions of homology. Much of the observed homology was localized to regions corresponding to consensus Ca2+-binding of E-F hand domains. Southern blot analysis suggested that the Dictyostelium genome contains a single gene encoding the EMLC. Examination of the pattern of EMLC mRNA expression showed that a significant increase in EMLC message levels occurred during the first few hours of development, coinciding with increased actin expression and immediately preceding the period of maximal chemotactic activity.

Amino Acid Sequence of Trocarin, a Prothrombin Activator FromTropidechis carinatus Venom: Its Structural Similarity to Coagulation Factor Xa

Blood ◽  
1999 ◽  
Vol 94 (2) ◽  
pp. 621-631 ◽  
Author(s):  
Jeremiah S. Joseph ◽  
Maxey C.M. Chung ◽  
Kandiah Jeyaseelan ◽  
R. Manjunatha Kini
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Blood Coagulation ◽  
Snake Venom ◽  
Heavy Chain ◽  
Light Chain ◽  
Critical Role ◽  
Factor Xa ◽  
Coagulation Factor ◽  
Group Ii

Abstract Among snake venom procoagulant proteins, group II prothrombin activators are functionally similar to blood coagulation factor Xa. We have purified and partially characterized the enzymatic properties of trocarin, the group II prothrombin activator from the venom of the Australian elapid, Tropidechis carinatus (rough-scaled snake). Prothrombin activation by trocarin is enhanced by Ca2+, phospholipids, and factor Va, similar to that by factor Xa. However, its amidolytic activity on peptide substrate S-2222 is significantly lower. We have determined the complete amino acid sequence of trocarin. It is a 46,515-Dalton glycoprotein highly homologous to factor Xa and shares the same domain architecture. The light chain possesses an N-terminal Gla domain containing 11 γ-carboxyglutamic acid residues, followed by two epidermal growth factor (EGF)-like domains; the heavy chain is a serine proteinase. Both chains are likely glycosylated: the light chain at Ser 52 and the heavy chain at Asn 45. Unlike other types of venom procoagulants, trocarin is the first true structural homologue of a coagulation factor. It clots snake plasma and thus may be similar, if not identical, to snake blood coagulation factor Xa. Unlike blood factor Xa, it is expressed in high quantities and in a nonhepatic tissue, making snake venom the richest source of factor Xa-like proteins. It induces cyanosis and death in mice at 1 mg/kg body weight. Thus, trocarin acts as a toxin in venom and a similar, if not identical, protein plays a critical role in hemostasis.

scholarly journals The complete amino acid sequence of human complement factor H

1988 ◽  
Vol 249 (2) ◽  
pp. 593-602 ◽  
Author(s):  
J Ripoche ◽  
A J Day ◽  
T J R Harris ◽  
R B Sim
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Repeat Unit ◽  
Regulatory Protein ◽  
Factor H ◽  
Cdna Clones ◽  
Complement Factor ◽  
Human Complement ◽  
Complete Amino Acid Sequence

The complete amino acid sequence of the human complement system regulatory protein, factor H, has been derived from sequencing three overlapping cDNA clones. The sequence consists of 1213 amino acids arranged in 20 homologous units, each about 60 amino acids long, and an 18-residue leader sequence. The 60-amino-acid-long repetitive units are homologous with those found in a large number of other complement and non-complement proteins. Two basic C-terminal residues, deduced from the cDNA sequence, are absent from factor H isolated from outdated plasma. A tyrosine/histidine polymorphism was observed within the seventh homologous repeat unit of factor H. This is likely to represent a difference between the two major allelic variants of factor H. The nature of the cDNA clones indicates that there is likely to be an alternative splicing mechanism, resulting in the formation of at least two species of factor H mRNA.

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