Updates of the HbVar database of human hemoglobin variants and thalassemia mutations

Abstract We compared the performance of two highly resolving methods, chromatofocusing (CRF) and isoelectric focusing in immobilized pH gradients (IPGF), for the separation of human hemoglobin variants. Lysates containing 13 different hemoglobins, including variants of clinical and geographical importance, and four electrophoretically "silent" variants (Hb Brockton, Hb Cheverly, Hb Köln, and Hb Waco) were analyzed. Both techniques showed a good intrarun precision (CV = 0.87% for CRF, 0.27% for IPGF) and high and similar resolving power (0.010 pH units, with the pH gradients used in this work). The use of an ultranarrow IPGF range (pH 7.15-7.35; pH gradient = 0.019 pH/cm) allowed the resolution between Hb Brockton, Hb Köln, and Hb A. In some cases (Hb D-Los Angeles, Hb F, Hb Waco), the variants were separated from Hb A in different orders, depending on which technique was used, probably because of the different analytical principles of the two methods. As a second-level test, both procedures are informative for characterization of human hemoglobin variants.

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Structural analysis of human hemoglobin variants by mass spectrometry

International Journal of Mass Spectrometry and Ion Physics ◽

10.1016/0020-7381(83)87065-x ◽

1983 ◽

Vol 48 ◽

pp. 209-212 ◽

Cited By ~ 19

Author(s):

Y. Wada ◽

A. Hayashi ◽

T. Fujita ◽

T. Matsuo ◽

I. Katakuse ◽

...

Keyword(s):

Mass Spectrometry ◽

Structural Analysis ◽

Human Hemoglobin ◽

Hemoglobin Variants

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Application of capillary isoelectric focusing with absorption imaging detection to the quantitative determination of human hemoglobin variants

Electrophoresis ◽

10.1002/elps.11501601107 ◽

1995 ◽

Vol 16 (1) ◽

pp. 670-673 ◽

Cited By ~ 35

Author(s):

Jiaqi Wu ◽

Janusz Pawliszyn

Keyword(s):

Quantitative Determination ◽

Isoelectric Focusing ◽

Human Hemoglobin ◽

Capillary Isoelectric Focusing ◽

Absorption Imaging ◽

Hemoglobin Variants ◽

Imaging Detection

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Are human hemoglobin variants distributed randomly among the positions?

Journal of Molecular Evolution ◽

10.1007/bf01653998 ◽

1973 ◽

Vol 2 (2-3) ◽

pp. 181-186 ◽

Cited By ~ 5

Author(s):

Walter M. Fitch

Keyword(s):

Human Hemoglobin ◽

Hemoglobin Variants

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HbVar: A relational database of human hemoglobin variants and thalassemia mutations at the globin gene server

Human Mutation ◽

10.1002/humu.10044 ◽

2002 ◽

Vol 19 (3) ◽

pp. 225-233 ◽

Cited By ~ 241

Author(s):

Ross C. Hardison ◽

David H.K. Chui ◽

Belinda Giardine ◽

Cathy Riemer ◽

George P. Patrinos ◽

...

Keyword(s):

Relational Database ◽

Globin Gene ◽

Human Hemoglobin ◽

Hemoglobin Variants

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THE STEPWISE MUTATION MODEL: AN EXPERIMENTAL EVALUATION UTILIZING HEMOGLOBIN VARIANTS

Genetics ◽

10.1093/genetics/94.1.185 ◽

1980 ◽

Vol 94 (1) ◽

pp. 185-201

Author(s):

Paul A Fuerst ◽

Robert E Ferrell

Keyword(s):

Amino Acid ◽

Electrophoretic Mobility ◽

Tertiary Structure ◽

Stepwise Mutation Model ◽

Starch Gel Electrophoresis ◽

Human Hemoglobin ◽

Net Charge ◽

Mutation Model ◽

Stepwise Mutation ◽

Hemoglobin Variants

ABSTRACT The stepwise mutation model of OHTA and KIMURA (1973) was proposed to explain patterns of genetic variability revealed by means of electrophoresis. The assumption that electrophoretic mobility was principally determined by unit changes in net molecular charge has been criticized by JOHNSON (1974, 1977). This assumption has been tested directly using hemoglobin. Twenty-seven human hemoglobin variants with known amino acid substitutions, and 26 nonhuman hemoglobins with known sequences were studied by starch gel electrophoresis. Of these hemoglobin% 60 to 70% had electrophoretic mobilities that could be predicted solely on the basis of net charge calculated from the amino acid composition alone, ignoring tertiary structure. Only four hemoglobins showed a mobility that was clearly different from an expected mobility calculated using only the net charge of the molecule. For the remaining 30% of hemoglobins studied, mobility was determined by a combination. of net charge and other unidentified components, probably reflecting changes in ionization of some amino acid residues as a result of small alterations in tertiary structure due to the amino acid substitution in the variant. For the nonhuman hemoglobins, the deviation of a sample from its expected mobility increased with increasing amino acid divergence from human hemoglobin A.—It is concluded that the net electrostatic charge of a molecule is the principal determinant of electrophoretic mobility under the conditions studied. However, because of the significant deviation from strict stepwise mobility detected for 30 to 40% of the variants studied, it is further concluded that the infinite-allele model of KIMURA and CROW (1964) or a "mixed model" such as that proposed by LI (1976) may be more appropriate than the stepwise mutation model for the analysis of much of the available electrophoretic data from natural populations.

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