Analysis of phenotypic manifestations of abnormal human hemoglobins

Phenotypic manifestations of abnormal human hemoglobins are discussed using the data of hematology, protein chemistry and molecular biology. On the basis of the presented analysis it is proposed to distinguish between phenotypic manifestations characterizing the primary molecular defect, i.e. properties of a mutant protein and their expression on the molecular and cellular levels; manifestations characterizing the equilibrium of the primary defect and compensatory potentialities of the organism; and finally the unbalanced state when the compensatory abilities of the organism are depleted. These different manifestations of the same defective gene reflect the most relevant peculiarities of mutant protein properties per se, expression of the properties in the living organism, the influence on the homeostatic system "mutant protein--organism" of the genetic and environmental factors both at the compensation state and at stress.     

A new strategy for structural identification of abnormal human hemoglobins

A new strategy for structural identification of abnormal human hemoglobins is proposed. It is based on micropreparative modification of electrophoretic separation of globins on Cellogel strips with subsequent quantitative isolation of a pure, desalted globin chain, in a form suitable for its subsequent structural investigation. Among the major advantages of the new strategy age possibility to use small blood samples (0.1-0.2 ml), short analysis time, relative simplicity and low cost.     

Identification of abnormal hemoglobins by isoelectric focusing

Using IEF on slabs of acrylamide gel was adapted for screening of abnormal Hemoglobins which are at the same level by electrophoresis on cellulose acetate strips. This method is fast, inexpensive and allowed the simultaneous analysis of 70 samples of whole blood. The characterization technique of IEF allowed us to distinguish some rare variants like Hb O Arab, HbD and T gamma in B 0-thalassemia.     

Micropreparative capillary zone electrophoresis of recombinant human interleukin-3

Micropreparative capillary zone electrophoresis of recombinant human interleukin-3 (rhIL-3) in untreated fused silica is presented. Results show that nanogram quantities of rhIL-3 can be collected off the capillary and then identified by amino acid sequencing and SDS-gel electrophoresis.     

Distribution of abnormal hemoglobins S and C in the Republic of Guinea]

Distribution of the HbS and HbC in the Guinean Republic was determined by the analysis of the excerption of 2213 inhabitants representing different ethnic groups of the country. It was found that the mean frequency of the HbS heterozygotes is 21.2 +/- 0.9% and of the HbC heterozygotes is 2.6 +/- 0.3%. Among major nationalities of the country the frequencies of the HbS heterozygotes make up 21.0 +/- 1.9% for the Fulbe, 22.2 +/- 1.6% for the Malinke and 26.5 +/- 1.6% for the Susu and frequencies of the HbC heterozygotes make up 3.0 +/- 0.8% for the Fulbe, 2.5 +/- 0.6% for the Malinke and 1.6 +/- 0.4% for the Susu. Relative viability of the HbS and HbC carriers for major Guinean nationalities is estimated. The relative fitnesses account is 1.05-1.13 for HbC heterozygotes and 1.07-1.16 for HbC heterozygotes.     

Plasma desorption mass spectrometry as a tool in characterization of abnormal proteins. Application to variant human hemoglobins.

We here report the application of plasma desorption mass spectrometry in combination with reversed-phase high-performance liquid chromatography and automatic Edman sequencing for the characterization of hemoglobin variants. By use of plasma desorption mass spectrometry to obtain molecular weight information of purified globin peptides it is possible to minimize the number of candidate positions for substitutions allowing an optimal use of automatic Edman degradation. Each variant can be characterized by using less than 200 micrograms of hemoglobin, corresponding to approximately 2 microliters of blood, as starting material. The outlined approach is considered to be very well suited for routine analysis of hemoglobins and other protein variants, natural as well as recombinant.