Variability in purified dysfunctional C1(-)-inhibitor proteins from patients with hereditary angioneurotic edema. Functional and analytical gel studies.

Donaldson, Virginia H.; Harrison, Richard A.; Rosen, Fred S.; Bing, David H.; Kindness, George; Canar, J; Wagner, Constance; Awad, Sameh

doi:10.1172/jci111664

Cited by 51 publications

(15 citation statements)

References 24 publications

(12 reference statements)

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“…as selective a loss of activity as did the mutant in this kindred (23,24). The best evidence that this mutation affects only target protease specificity and not the inhibitory mechanism is that, although it has lost activity against Clr, it has acquired inhibitory activity against trypsin (Fig.…”

Section: Methodsmentioning

confidence: 58%

Unique C1 inhibitor dysfunction in a kindred without angioedema. II. Identification of an Ala443-->Val substitution and functional analysis of the recombinant mutant protein.

Zahedi¹,

Bissler²,

Davis³

et al. 1995

J. Clin. Invest.

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We have determined the cause of an unusual Cl inhibitor abnormality in a large kindred. We previously found that half of serum Cl inhibitor molecules in affected kindred members are normal. The other half complexed with Cls but showed little complex formation with Clr. These molecules also appeared to be relatively resistant to digestion by trypsin. Taken together, the findings suggested that members of this kindred are heterozygous for an unusual Cl inhibitor mutation. Sequencing of genomic DNA from the kindred revealed that thymine has replaced cytosine in the codon for Ala443 (P2 residue) in one Cl inhibitor allele, resulting in substitution with a Val residue. To test the effect of this substitution, a mutant Cl inhibitor containing Ala"3--Val was constructed by site-directed mutagenesis and expressed in COS-1 cells. Both the Ala'3--Val mutant and the wild-type Cl inhibitor complexed completely with Cls, kallikrein, and coagulation Factor XIIa after incubation at 37°C for 60 min. In contrast, the mutant inhibitor failed to complex completely with Clr under the same conditions. Time course analysis showed that the ability of the mutant to complex with Cls is also impaired: although it complexed completely in 60 min, the rate of complex formation during a 0-60-min incubation was decreased compared with wild-type Cl inhibitor. The mutant inhibitor also formed a complex with trypsin, a serine protease that cleaves, and is not inhibited by, wild-type Cl inhibitor. The Ala"3-Val mutation therefore converts Cl inhibitor from a substrate to an inhibitor of trypsin. These studies emphasize the role of the P2 residue in the determination of target protease specificity. (J. Clin. Invest. 1995. 95:1299-1305

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Section: Methodsmentioning

confidence: 58%

Unique C1 inhibitor dysfunction in a kindred without angioedema. II. Identification of an Ala443-->Val substitution and functional analysis of the recombinant mutant protein.

Zahedi¹,

Bissler²,

Davis³

et al. 1995

J. Clin. Invest.

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“…Heterogeneity of the dysmorphic proteins has been demonstrated among different type-II HANE families (5). Studies ofin vitro cell cultures have shown that monocytes from patients with HANE synthesize -50% of normal CI-Inh compared with cells from normal subjects (6), which is in agreement with the autosomal dominant inheritance of the disorder (7).…”

Section: Introductionmentioning

confidence: 63%

Plasma levels of C1- inhibitor complexes and cleaved C1- inhibitor in patients with hereditary angioneurotic edema.

et al. 1990

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“…Genetic analysis of HANE indicates that the molecular genetic defects responsible for Cl INH deficiency are heterogeneous, as are those responsible for many other genetic diseases (1 [1][2][3][4][5][6][7][8][9][10][11][12][13][14]. Analysis of mutant proteins from type II HANE patients provided the first evidence for genetic heterogeneity: the proteins from different families differ in electrophoretic mobility, size, and function (6,15,16). Evaluation of the Cl INH gene by Southern blot analysis in both types of HANE has revealed RFLP in -10-15% of kindred (10).…”

Section: Introductionmentioning

confidence: 99%

Type I C1 inhibitor deficiency with a small messenger RNA resulting from deletion of one exon.

Ariga¹,

Igarashi²,

Ramesh³

et al. 1989

J. Clin. Invest.

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The molecular genetic basis of C1 inhibitor (Cl INH) deficiency in a patient with type I hereditary angioneurotic edema was studied. This patient was found to have an abnormally short C1 INH mRNA together with a normal message. Restriction fragment length polymorphism of the Cl INH gene was detected by Southern blot analysis of the patient's DNA after digestion with Pst I or Sac I, and hybridization with a full-length C1 INH cDNA. Hybridization of the same blot with three different fragments of the full-length cDNA suggested that exon VII and portions of both flanking introns were deleted in the C1 INH gene. Northern blot analysis of RNA from cultured monocytes, using a probe corresponding to exon VII, also indicated that the abnormal C1 INH mRNA had a deletion of these nucleotides. To confirm the hypothesis that the short C1 INH mRNA contained a deletion, the involved segment of the patient's C1 INH mRNA was amplified using the polymerase chain reaction (PCR). PCR amplification yielded two C1 INH DNA fragments of different lengths (380 and 160 bp). Southern blot and sequence analysis of both DNA fragments clearly revealed that the smaller 160-bp DNA was derived from the abnormal message and had a deletion of nucleotides corresponding to exon VII.

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Variability in purified dysfunctional C1(-)-inhibitor proteins from patients with hereditary angioneurotic edema. Functional and analytical gel studies.

Abstract: Introduction

Cited by 51 publications

References 24 publications

Unique C1 inhibitor dysfunction in a kindred without angioedema. II. Identification of an Ala443-->Val substitution and functional analysis of the recombinant mutant protein.

Unique C1 inhibitor dysfunction in a kindred without angioedema. II. Identification of an Ala443-->Val substitution and functional analysis of the recombinant mutant protein.

Plasma levels of C1- inhibitor complexes and cleaved C1- inhibitor in patients with hereditary angioneurotic edema.

Type I C1 inhibitor deficiency with a small messenger RNA resulting from deletion of one exon.

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