Hb G-Philadelphia, or [α68(E17)Asn → Lys], in North Sardinia: detection by isoelectric focusing and identification by HPLC of tryptic peptides

Manca, Laura; Demuro, P; Masala, Bruno Lucio

doi:10.1016/0009-8981(88)90067-8

Cited by 19 publications

(6 citation statements)

References 19 publications

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“…Isoelectric focusing (IEF) on thin‐layer 5% polyacrylamide slab gels (pH range 3.5–10.0) was performed as described by Manca et al . [19] and Masala & Manca [20]. Gels were stained for proteins using 0.1% bromphenol blue solution.…”

Section: Methodsmentioning

confidence: 99%

The hemoglobin system of the brown moray Gymnothorax unicolor

Tamburrini

Verde

Olianas

et al. 2001

European Journal of Biochemistry

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The Gymnothorax unicolor hemoglobin system is characterized by two components, called cathodic and anodic on the basis of their isoelectric point, which were separated by ion-exchange chromatography. The oxygen-binding properties of the purified components were studied in the absence and presence of chloride and/or GTP or ATP in the pH range 6.5-8.0. Stripped cathodic hemoglobin showed a small reverse Bohr effect, high oxygen affinity, and low co-operativity; the addition of chloride only caused a small decrease in oxygen affinity. In the presence of GTP or ATP, the oxygen affinity was dramatically reduced, the co-operativity increased, and the reverse Bohr effect abolished. Stripped anodic hemoglobin is characterized by both low oxygen affinity and co-operativity, and displayed a normal Bohr effect; the addition of chloride increased co-operativity, whereas ATP and GTP significantly modulated oxygen affinity at acidic pH values, enhancing the Bohr effect and giving rise to the Root effect. The complete amino-acid sequences of the alpha and beta chains of both hemoglobins were established; the molecular basis of the functional properties of the hemoglobins is discussed in the light of the primary structure and compared with those of other fish hemoglobins.

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Section: Methodsmentioning

confidence: 99%

The hemoglobin system of the brown moray Gymnothorax unicolor

Tamburrini

Verde

Olianas

et al. 2001

European Journal of Biochemistry

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“…The purity of the pig chains and hybrid haemoglobins was checked by alkaline PAGE, isoelectric focusing on thin-layer 5 % polyacrylamide gel (Ampholine pH range 6-8 ; Pharmacia LKB Biotechnology) [15,16] and acid PAGE in 8 M urea and Triton X-100 under denaturing conditions (AUT\PAGE) [17].…”

Section: Gel Electrophoresis Of Dissociated Globin Chainsmentioning

confidence: 99%

Circe's haemoglobins, pig–human hybrids: functional characterization and structural considerations

Sanna

Giardina

Pellegrini

et al. 1998

Biochemical Journal

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We report the isolation and the functional characterization of alpha and beta chains from pig (Sus scropha domesticus) haemoglobin, as well as of the pig-human hybrid haemoglobins, alpha2(h)beta2(p) and alpha2(p)beta2(h) (i.e. Circe's haemoglobins), obtained by mixing the purified alpha and beta pig chains respectively with the corresponding partner human chains. Their functional properties have been compared with those of both parental haemoglobins in order to obtain information on the role of the different subunits and of their inter-relationships, both at the structural and functional levels. The results indicate that the functional properties of both hybrids are closer to those of the parental haemoglobin that provides the beta chains, confirming the major role of the beta chains in determining the oxygen affinity and the modulation mechanisms of the tetrameric molecule. This is supported by the thermodynamic properties, since the very low DeltaH of oxygen binding that characterizes pig haemoglobin and the alpha2(h)beta2(p) hybrid haemoglobin may be taken as the reflection of specific structural properties of pig beta chain.

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“…In two subjects with 37% level of the variant, the C→G mutation was indeed on the α 2 gene, while the hybrid − α 3.7 gene was present in trans . In a previous study (9), we assumed that the trimodal distribution of the level of the variant among Sardinians (with modes centering on 28, 36, and 42%, respectively) was because of the occurrence of the mutation both on a double and on a single locus chromosome, and that the linkage with α ‐thal determinants could be partially responsible for its rather high frequency. In fact, the − α 3.7 thal gene reaches considerably high incidences in Sardinia, ranging from 23% in the North and 33% in the South (10).…”

Section: Discussionmentioning

confidence: 99%

“…We described previously that the Hb G is rather common in north Sardinia, its prevalence being approximately 1 in 1600 (9, 10). Its level, as percentage of total Hb, has a trimodal distribution: 27.8 ± 2.3, 36.3 ± 2.4, 42.3 ± 1.5.…”

mentioning

confidence: 99%

The C→G transition in the α2‐globin gene of a normal αα‐chromosome is responsible for the Hb G‐Philadelphia variant in Sardinians

Masala

Musino

Pirastru

et al. 2004

European J of Haematology

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Sequencing of alpha-globin genes of 18 Sardinian heterozygotes for the Hb G-Philadelphia [alpha 68(E17)Asn-->Lys] variant, with four active alpha genes and circulating level of the variant of about 27%, showed the AAC-->AAG change at codon 68 of the alpha 2-globin gene (alpha(G)alpha/alpha alpha). Two heterozygotes with level of about 37% were the carriers of the same mutation on the same alpha 2 gene, and of the alpha 2 alpha 1 hybrid gene, because of the 3.7-kb deletion, in trans (alpha(G)alpha/-alpha(3.7)). In Black people, the same C-->G mutation occurs on the hybrid gene (-alpha(G)3.7), whereas in Caucasians the Lys for Asn change is because of the C-->A transversion occurring on the alpha 2 gene of a normal alpha alpha arrangement. The identification of the C-->G mutation on the normal alpha alpha chromosome points to an undescribed genotype for this rather common variant, which is probably because of the high rate of recombination between the duplicated alpha-globin genes.

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Hb G-Philadelphia, or [α68(E17)Asn → Lys], in North Sardinia: detection by isoelectric focusing and identification by HPLC of tryptic peptides

Cited by 19 publications

References 19 publications

The hemoglobin system of the brown moray Gymnothorax unicolor

The hemoglobin system of the brown moray Gymnothorax unicolor

Circe's haemoglobins, pig–human hybrids: functional characterization and structural considerations

The C→G transition in the α2‐globin gene of a normal αα‐chromosome is responsible for the Hb G‐Philadelphia variant in Sardinians

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