scholarly journals Immunochemical Quantitation of Proteins in Single Cells

Blood ◽  
1968 ◽  
Vol 32 (5) ◽  
pp. 796-810 ◽  
Author(s):  
DAVID GITLIN ◽  
TERUO SASAKI ◽  
PEKKA VUOPIO
Keyword(s):  
Carbonic Anhydrase ◽  
Single Cells ◽  
Immunochemical Method ◽  
Carbonic Anhydrase B ◽  
Immunochemical Quantitation ◽  
Unusual Distribution ◽  
Chain Synthesis ◽  
Β Chain ◽  
Individual Erythrocytes

Abstract An immunochemical method for the quantitative determination of specific soluble proteins in individual erythrocytes has been described. Application of the method revealed: 1. From 0.5 to 0.9 per cent of the normal adult erythrocytes studied contained small amounts of γ-chains, from 2.5 to 12.5 µµg. as hemoglobin per cell. 2. In cord blood of normal infants of 38 weeks’ gestation, significant numbers of erythrocytes were found which contained either γ-chains or β-chains or both, independently of carbonic anhydrase B. 3. Once initiated within a given erythrocyte, β-chain synthesis in that cell rapidly approached adult rates. Derepression of carbonic anhydrase B synthesis was independent of derepression of hemoglobin synthesis, and in fetal cells which contained carbonic anhydrase B the amount found was well below that of most adult erythrocytes. 4. In 4 patients homozygous for hemoglobin S and in 2 persons heterozygous for thalassemia, an increase in F hemoglobin was associated with an increase in γ-chain content of individual erythrocytes as well as an increase in the number of erythrocytes containing γ-chains; in 2 patients homozygous for thalassemia, an unusual distribution of cells with increased γ-chain content was observed.

Mutant Fetal Hemoglobin Causing Cyanosis in a Newborn

PEDIATRICS ◽  
1989 ◽  
Vol 83 (5) ◽  
pp. 734-736
Author(s):  
John R. Priest ◽  
Jan Watterson ◽  
Richard T. Jones ◽  
Anne E. Faassen ◽  
Bo E. Hedlund
Keyword(s):  
Amino Acid ◽  
Amino Acid Substitution ◽  
Clinical Evidence ◽  
Fetal Hemoglobin ◽  
Single Amino Acid Substitution ◽  
Single Amino Acid ◽  
Globin Chain ◽  
Chain Synthesis ◽  
A Site ◽  
Β Chain

A well but cyanotic newborn was found to have a mutant γ-globin chain, leading to a functionally abnormal fetal hemoglobin. A single amino acid substitution was found in a site consistent with known adult M hemoglobins. This patient showed no clinical evidence of cyanosis at 5 weeks of age as γ-chain synthesis was replaced by β-chain synthesis. A sibling born 20 months later was also cyanotic and the same mutant hemoglobin was found.

Two slow stages in refolding of bovine carbonic anhydrase B are due to proline isomerization

1990 ◽  
Vol 213 (3) ◽  
pp. 561-568 ◽  
Author(s):  
G.V. Semisotnov ◽  
V.N. Uversky ◽  
I.V. Sokolovsky ◽  
A.M. Gutin ◽  
O.I. Razgulyaev ◽  
...  
Keyword(s):  
Carbonic Anhydrase ◽  
Bovine Carbonic Anhydrase ◽  
Proline Isomerization ◽  
Carbonic Anhydrase B

scholarly journals Polymorphisms in Brucella Carbonic anhydrase II mediate CO2 dependence and fitness in vivo

2019 ◽  
Author(s):  
JM García-Lobo ◽  
Y Ortiz ◽  
C González-Riancho ◽  
A Seoane ◽  
B Arellano-Reynoso ◽  
...  
Keyword(s):  
Carbonic Anhydrase ◽  
Primary Cultures ◽  
Low Frequency ◽  
Carbonic Anhydrase Ii ◽  
Carbonic Anhydrases ◽  
Significant Difference ◽  
Dependent Strain

AbstractSome Brucella isolates are known to require an increased concentration of CO2 for growth, especially in the case of primary cultures obtained directly from infected animals. Moreover, the different Brucella species and biovars show a characteristic pattern of CO2 requirement, and this trait has been included among the routine typing tests used for species and biovar differentiation. By comparing the differences in gene content among different CO2-dependent and CO2-independent Brucella strains we have confirmed that carbonic anhydrase II (CA II), is the enzyme responsible for this phenotype in all the Brucella strains tested. Brucella species contain two carbonic anhydrases of the β family, CA I and CA II; genetic polymorphisms exist for both of them in different isolates, but only those putatively affecting the activity of CA II correlate with the CO2 requirement of the corresponding isolate. Analysis of these polymorphisms does not allow the determination of CA I functionality, while the polymorphisms in CA II consist of small deletions that cause a frameshift that changes the C-terminus of the protein, probably affecting its dimerization status, essential for the activity.CO2-independent mutants arise easily in vitro, although with a low frequency ranging from 10−6 to 10−10 depending on the strain. These mutants carry compensatory mutations that produce a full length CA II. At the same time, no change was observed in the sequence coding for CA I. A competitive index assay designed to evaluate the fitness of a CO2-dependent strain compared to its corresponding CO2-independent strain revealed that while there is no significant difference when the bacteria are grown in culture plates, growth in vivo in a mouse model of infection provides a significant advantage to the CO2-dependent strain. This could explain why some Brucella isolates are CO2-dependent in primary isolation. The polymorphism described here also allows the in silico determination of the CO2 requirement status of any Brucella strain.

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