The alpha chains of goat hemoglobins: Old and new variants in native Apulian breeds

Pieragostini, E.; Rullo, Rosario; Scaloni, Andrea; Bramante, G.; Luccia, Aldo Di

doi:10.1016/j.cbpc.2005.05.005

Cited by 10 publications

(10 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On this basis, no functional consequences of the p.Ala27Thr substitution can be predicted according to known site mutation determinants (Storz and Moriyama 2008;Weber and Campbell 2011). The same amino acid replacement at the same position has already been detected in α-chain globin variants from goat breeds present in southern Italy (Puglia and Sardegna) (Pieragostini et al 2005;Pirastru et al 2009) and from human families in Caserta (Campania) (Lacerra et al 2004). Either in Agerolese bovine, Apulian/Sardinian goat, and human Casertian α-globin variants, this substitution involves a CpG dinucleotide that is considered as a mutation hotspot (Perutz 1990).…”

Section: Resultsmentioning

confidence: 88%

“…Blood lysates were analyzed by polyacrylamide isoelectrofocusing (PAGIF) over a 6.6-7.6 pH gradient (Scaloni et al 1998). They were also analyzed by acid-urea triton X-100 polyacrylamide gel electrophoresis (AUT-PAGE), as previously reported (Pieragostini et al 2005). Blood lysates were also diluted (1 : 100) in 10% acetonitrile (ACN), 0.1% trifuoroacetic acid (TFA), and analyzed by matrix assisted laser desorption ionization-time of flightmass spectrometry (MALDI-TOF-MS) using a UltrafleXtreme TM instrument (Bruker Daltonics, Bremen, Germany).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Bovine hemoglobin polymorphism: a novel alpha-globin variant identified in the Agerolese breed from southern Italy

et al. 2015

View full text Add to dashboard Cite

This study describes the results of molecular screening in the Agerolese cattle breed from southern Italy, which constitutes an economical source and contributes to the regional biodiversity. Integrated electrophoretic, chromatographic, mass spectrometric, and DNA sequencing procedures were used to qualitatively and quantitatively evaluate globin chain polymorphism in a population of animals from different farms. In addition to α-, β A -, and β B -globin-bearing animal phenotypes, our data demonstrate the occurrence of individuals characterized by the presence of a novel α-globin variant, whose primary structure differs from the normal counterpart for the p.Ala27Thr substitution. The SNP c.79G>A located at the exon 1 of the HBA2 gene is responsible for this amino acid change and it represents a third example of α-globin gene polymorphism observed in Bos taurus. This phenomenon was ascertained only in two Mediterranean breeds from southern Italy and highlights the uncommon occurrence of α-globin variants in this geographical region, similarly to what has already been described in goat and human.

show abstract

Section: Resultsmentioning

confidence: 88%

Section: Methodsmentioning

confidence: 99%

Bovine hemoglobin polymorphism: a novel alpha-globin variant identified in the Agerolese breed from southern Italy

et al. 2015

View full text Add to dashboard Cite

show abstract

“…2003; Iorio et al. 2004; Pieragostini et al. 2005) and with the guidelines for gene nomenclature in ruminants (Andresen et al.…”

Section: Resultsmentioning

confidence: 99%

Functional effect of haemoglobin polymorphism on the haematological pattern of Gentile di Puglia sheep

Pieragostini

Rubino

Bramante

et al. 2006

J Animal Breeding Genetics

View full text Add to dashboard Cite

The relationship between haemoglobin polymorphism and haematological pattern were studied in Gentile di Puglia, the Apulia Merino native breed. In a sample of 292 individuals, on five different farms, alpha- and beta-globin genetic polymorphism was investigated. A remarkable polymorphism was detected especially at alpha-globin gene where the variations concerned both the quality of the gene product and gene expression. Triplicated and quadruplicated alpha-globin genes were observed in 8.6% and 1.2% of the alpha-haplotypes respectively. At beta-globin (HBB) locus, predominance of HBB(B) allele was found in all flocks, while HBB(A) overall frequency was around 11.2%. Moreover, the effect of genotypes at globin systems on haematological data was evaluated on 289 animals. A significant effect was detected for HBB locus on haematocrit (HCT) and mean corpuscular volume (MCV), with decreasing HCT and MCV for decreasing number of HBB(A) alleles in the genotype. The opposite trend was observed for mean corpuscular haemoglobin concentration. The advances presented by this paper give reasons for inferring that BB genotypes can be expected to be more resilient to climatic stress and to local parasites.

show abstract

“…To assess whether the N-terminal methionine residues were properly cleaved, N-terminal peptide sequences of the α- and β-chain subunits were determined by means of Edman degradation. To prepare samples for peptide sequencing, the α- and β-globin monomers were separated by means of Acid-Urea-Triton (AUT) polyacrylamide gel electrophoresis [33], and were then transferred to PVDF membrane. The efficacy of the MAP enzyme in cleaving the N-terminal methionine residues was optimized by systematically altering the induction temperature, the induction time, and E. coli expression strain.…”

Section: Methodsmentioning

confidence: 99%

Expression and Purification of Recombinant Hemoglobin in Escherichia coli

et al. 2011

View full text Add to dashboard Cite

BackgroundRecombinant DNA technologies have played a pivotal role in the elucidation of structure-function relationships in hemoglobin (Hb) and other globin proteins. Here we describe the development of a plasmid expression system to synthesize recombinant Hbs in Escherichia coli, and we describe a protocol for expressing Hbs with low intrinsic solubilities. Since the α- and β-chain Hbs of different species span a broad range of solubilities, experimental protocols that have been optimized for expressing recombinant human HbA may often prove unsuitable for the recombinant expression of wildtype and mutant Hbs of other species.Methodology/Principal FindingsAs a test case for our expression system, we produced recombinant Hbs of the deer mouse (Peromyscus maniculatus), a species that has been the subject of research on mechanisms of Hb adaptation to hypoxia. By experimentally assessing the combined effects of induction temperature, induction time and E. coli expression strain on the solubility of recombinant deer mouse Hbs, we identified combinations of expression conditions that greatly enhanced the yield of recombinant protein and which also increased the efficiency of post-translational modifications.Conclusion/SignificanceOur protocol should prove useful for the experimental study of recombinant Hbs in many non-human animals. One of the chief advantages of our protocol is that we can express soluble recombinant Hb without co-expressing molecular chaperones, and without the need for additional reconstitution or heme-incorporation steps. Moreover, our plasmid construct contains a combination of unique restriction sites that allows us to produce recombinant Hbs with different α- and β-chain subunit combinations by means of cassette mutagenesis.

show abstract

The alpha chains of goat hemoglobins: Old and new variants in native Apulian breeds

Cited by 10 publications

References 35 publications

Bovine hemoglobin polymorphism: a novel alpha-globin variant identified in the Agerolese breed from southern Italy

Bovine hemoglobin polymorphism: a novel alpha-globin variant identified in the Agerolese breed from southern Italy

Functional effect of haemoglobin polymorphism on the haematological pattern of Gentile di Puglia sheep

Expression and Purification of Recombinant Hemoglobin in Escherichia coli

Contact Info

Product

Resources

About