Two novel mutations responsible for hereditary type I protein C deficiency: Characterization by denaturing gradient gel electrophoresis

1992 ◽  
Vol 1 (6) ◽  
pp. 491-500 ◽  
Author(s):  
S. Gandrille ◽  
M. Vidaud ◽  
M. Aiach ◽  
M. Alhenc-Gelas ◽  
A. M. Fischer ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Protein C ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Type I ◽  
Novel Mutations ◽  
Protein C Deficiency ◽  
Gradient Gel Electrophoresis

scholarly journals Five novel mutations located in exons III and IX of the protein C gene in patients presenting with defective protein C anticoagulant activity

Blood ◽  
1993 ◽  
Vol 82 (1) ◽  
pp. 159-168 ◽  
Author(s):  
S Gandrille ◽  
M Alhenc-Gelas ◽  
P Gaussem ◽  
MF Aillaud ◽  
E Dupuy ◽  
...  
Keyword(s):  
Amino Acids ◽  
Protein C ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Anticoagulant Activity ◽  
Direct Sequencing ◽  
Catalytic Triad ◽  
Activity Levels ◽  
Novel Mutations ◽  
Protein C Deficiency

We describe five families presenting with type II hereditary protein C deficiency characterized by normal antigen and amidolytic activity levels but low anticoagulant activity. All the exons and intron/exon junctions of the protein C gene were studied using a strategy combining amplification by the polymerase chain reaction (PCR), denaturing gradient gel electrophoresis of the amplified fragments, and direct sequencing of fragments displaying altered melting behavior. We detected five novel mutations. Three were located in the C-terminal part of the propeptide encoded by exon III: Arginine (Arg)-5 to tryptophan (Trp), Arg-1 to histidine (His), and Arg-1 to cysteine (Cys) mutations. The two others, located in exon IX, affected Arg 229 and serine (Ser) 252, which were respectively replaced by glutamine (Gln) and asparagine (Asn). DNA studies of the other exons from affected individuals showed no other abnormalities. These novel mutations provide further insight into the importance of the affected amino acids located close to the active site, near Asp 257, one of the three amino acids of the catalytic triad. The low anticoagulant activity of the abnormal protein C indicated that Arg 229 and Ser 252 play a key role during the interaction between protein C and its cofactor protein S, phospholipids, or factors Va and VIIIa. The Arg-1 to Cys mutation led to the dimerization of protein C with another plasmatic component, as evidenced by the presence in the plasma of a high molecular weight form of protein C that disappeared after reduction. No molecular mass abnormalities were observed in heavy and light chains of all other protein C mutants. In the five families explored, 9 (64%) of the 14 subjects bearing the mutations reported thrombotic events. This suggests that the protein C amino acids affected by the mutations are very important for the in vivo expression of the antithrombotic properties of protein C.

Five Novel Mutations of the Protein S Active Gene (PROS 1) in 8 Norman Families

1996 ◽  
Vol 75 (03) ◽  
pp. 437-444 ◽  
Author(s):  
Jérôme Duchemin ◽  
Jeanne-Yvonne Borg ◽  
Delphine Borgel ◽  
Marc Vasse ◽  
Hervé Lévèque ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Molecular Basis ◽  
Protein S ◽  
Type I ◽  
Factor V ◽  
Novel Mutations ◽  
Nucleotide Deletion ◽  
Active Gene ◽  
Sequence Variations ◽  
Gradient Gel Electrophoresis

SummaryTo further elucidate the molecular basis for hereditary thrombophilia, we screened the protein S active gene in 11 families with type I deficiency, using a strategy based on denaturating gradient gel electrophoresis (DGGE) of all the coding sequences. Fragments with an abnormal DGGE pattern were sequenced, and 5 novel mutations were identified in 8 families. The mutations were a 7-nucleotide deletion in exon II, a 4-nucleotide deletion in exon III, a T insertion in exon VII, a C to T transition transforming Leu 259 into Pro and a T to C transition transforming Cys 625 into Arg in 4 families. These mutations were the only sequence variations found in the propositus’ gene exons and co-segregated with the plasma phenotype. A total of 28 members of these 8 families were heterozygous for one of the 5 mutations. Twenty-four (58,5%) of the 41 deficient subjects over 18 years of age had clinical thrombophilia, whereas the 13 subjects under 18 were asymptomatic. Of the 28 subjects, 6 (21,5%) were also found to bear the factor V Arg 506 Gin mutation.

scholarly journals Analysis of Methane Monooxygenase Genes in Mono Lake Suggests That Increased Methane Oxidation Activity May Correlate with a Change in Methanotroph Community Structure

2005 ◽  
Vol 71 (10) ◽  
pp. 6458-6462 ◽  
Author(s):  
Ju-Ling Lin ◽  
Samantha B. Joye ◽  
Johannes C. M. Scholten ◽  
Hendrik Schäfer ◽  
Ian R. McDonald ◽  
...  
Keyword(s):  
Methane Oxidation ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Mono Lake ◽  
Hypersaline Lake ◽  
Type I ◽  
Type Ii ◽  
Oxidation Activity ◽  
Oxidation Rates ◽  
Gradient Gel Electrophoresis

ABSTRACT Mono Lake is an alkaline hypersaline lake that supports high methane oxidation rates. Retrieved pmoA sequences showed a broad diversity of aerobic methane oxidizers including the type I methanotrophs Methylobacter (the dominant genus), Methylomicrobium, and Methylothermus, and the type II methanotroph Methylocystis. Stratification of Mono Lake resulted in variation of aerobic methane oxidation rates with depth. Methanotroph diversity as determined by analysis of pmoA using new denaturing gradient gel electrophoresis primers suggested that variations in methane oxidation activity may correlate with changes in methanotroph community composition.

Thirty-three Novel Mutations in the Protein C Gene

2000 ◽  
Vol 83 (01) ◽  
pp. 86-92 ◽  
Author(s):  
Sophie Gandrille ◽  
Marie-Laurence Aubry ◽  
Martine Aiach ◽  
Martine Alhenc-Gelas ◽  
Keyword(s):  
Protein C ◽  
Sequence Variation ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Gene Mutations ◽  
Promoter Polymorphisms ◽  
Scanning Strategy ◽  
Novel Mutations ◽  
Sequence Variations ◽  
Gradient Gel Electrophoresis ◽  
Proc Gene

SummaryWe analyzed the protein C gene (PROC) with the denaturing gradient gel electrophoresis (DGGE) scanning strategy in a series of 129 patients with suspected protein C (PC) deficiency (93 with low plasma PC levels and 36 with borderline level). At least one sequence variation was found in 104 of the 129 patients. Thirty-nine sequence variations (found in 72 patients) were already reported detrimental mutations. Thirty-three were novel sequence variations, of which 19 (found in 25 patients) were probably detrimental. Five novel mutations (A1T, R9H, S11R, S12R and K193Q) were associated with qualitative plasma PC deficiency, suggesting or confirming the functional importance of amino acids at these positions. This strategy confirmed the diagnosis of inherited PC deficiency in 79/93 (84.9%) patients with low plasma PC levels and 14/36 (38.8%) patients with borderline values. In order to explain abnormal PC levels observed in patients who did not carry detrimental mutations, screening for the -1654C/T and -1641A/G PROC promoter polymorphisms known to influence plasma PC concentrations was performed. The frequency of the CG allele associated with lower PC concentrations was slightly but not significantly lower in 82 heterozygotes for detrimental PROC gene mutations than in 36 patients with no identified detrimental mutations.

Novel mutations identified using a comprehensive CCR5-denaturing gradient gel electrophoresis assay

AIDS ◽  
2001 ◽  
Vol 15 (2) ◽  
pp. 171-177 ◽  
Author(s):  
Desiree C. Petersen ◽  
Maritha J. Kotze ◽  
Michele D. Zeier ◽  
Ashraf Grimwood ◽  
Deon Pretorius ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Novel Mutations ◽  
Gradient Gel Electrophoresis

Novel mutations and SNPs identified inCCR2 using a new comprehensive denaturing gradient gel electrophoresis assay

2002 ◽  
Vol 20 (4) ◽  
pp. 253-259 ◽  
Author(s):  
Desiree C. Petersen ◽  
Annette Laten ◽  
Michele D. Zeier ◽  
Ashraf Grimwood ◽  
Estrelita Janse van Rensburg ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Novel Mutations ◽  
Gradient Gel Electrophoresis

The Frequency of Type I Heterozygous Protein S and Protein C Deficiency in 141 Unrelated Young Patients with Venous Thrombosis

1988 ◽  
Vol 59 (01) ◽  
pp. 018-022 ◽  
Author(s):  
C L Gladson ◽  
I Scharrer ◽  
V Hach ◽  
K H Beck ◽  
J H Griffin
Keyword(s):  
Venous Thrombosis ◽  
Protein C ◽  
Antithrombin Iii ◽  
Young Patients ◽  
Protein S ◽  
Type I ◽  
Protein S Deficiency ◽  
Protein C Deficiency ◽  
Low Protein ◽  
S Deficiency

SummaryThe frequency of heterozygous protein C and protein S deficiency, detected by measuring total plasma antigen, in a group (n = 141) of young unrelated patients (<45 years old) with venous thrombotic disease was studied and compared to that of antithrombin III, fibrinogen, and plasminogen deficiencies. Among 91 patients not receiving oral anticoagulants, six had low protein S antigen levels and one had a low protein C antigen level. Among 50 patients receiving oral anticoagulant therapy, abnormally low ratios of protein S or C to other vitamin K-dependent factors were presented by one patient for protein S and five for protein C. Thus, heterozygous Type I protein S deficiency appeared in seven of 141 patients (5%) and heterozygous Type I protein C deficiency in six of 141 patients (4%). Eleven of thirteen deficient patients had recurrent venous thrombosis. In this group of 141 patients, 1% had an identifiable fibrinogen abnormality, 2% a plasminogen abnormality, and 3% an antithrombin III deficiency. Thus, among the known plasma protein deficiencies associated with venous thrombosis, protein S and protein C. deficiencies (9%) emerge as the leading identifiable associated abnormalities.

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