The Amino Acid Sequences of the α- and β-Globin Chains of Hemoglobin from the Aldabra Giant Tortoises, Geochelone gigantea

Shishikura, Fumio; Takami, Kazutoshi

doi:10.2108/zsj.18.515

Cited by 8 publications

(8 citation statements)

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“…However, the author can extract useful information that the two giant tortoises including the small Chaco tortoises separated after the breakup of Gondwana. Table 3 shows that the separation times of the three species occurred approximately 12-20 mya, which supports grossly our previous reports [5][6][7] as well as the mtDNA studies of Caccone et al 14,18) . Fig.…”

Section: Molecular Relationships and Divergence Timessupporting

confidence: 89%

“…On the contrary, the author has established the molecular phylogeny of the three populations of Geochelone (one from Africa and two from South America) based on the primary structures of 53 subunits of amniote globins, including those of seven species of reptiles, three species of birds, and humans 7) . The divergence time of the Galapagos giant tortoises and the Aldabra giant tortoises was estimated to be 15-21 mya on the basis of globin genealogy [5][6][7] . Even though their present habitats are separated by great distance, it was shown that both giant tortoise species diverged during the early Oligocene to the late Miocene epochs, which is supported by recent studies of the two groups 19,20) .…”

Section: Molecular Relationships and Divergence Timesmentioning

confidence: 99%

“…Here, the author aims to provide scientific information on the origin and the divergence time of the two giant tortoise species, including a close relative to the Galapagos giant tortoises, the Chaco tortoise (Geochelone chilensis; synonym: Chelonoidis chilensis). To analyze intra-genus relationships, the author first constructed a molecular tree based on the primary structure of the globin subunits α A , α D , and β [5][6][7] . Both giant tortoise species were shown to diverge around 15-21 mya 6,7) , which represents a significant delay following the break-up of Gondwana, which separated into Africa and South America during the Cretaceous period 65-135 mya 8,9) .…”

Section: Introductionmentioning

confidence: 99%

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Globin-intron Sequences Imply Molecular Relationships Between the Galapagos Giant Tortoise and the Aldabra Giant Tortoise

Shishikura

2013

Nichidai Igaku Zasshi

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The author previously established a molecular tree based on the cDNA-derived primary structures of globins in two giant tortoises, Geochelone nigra and G. gigantea. The divergence time was estimated to be 15-21 million years ago. In the present study, the author reexamined the divergence time of these two species using another source of genetic information-globin-introns-including those from the Chaco tortoise (G. chilensis), a close relative of G. nigra. The previously determined divergence time was supported by the findings of this intron study. However, the inter-relationships based on the intron nucleotide sequences of the globins from the three Geochelone species remain controversial, because it is difficult to determine which of the three is the ancestral species. In addition, the nucleotide sequences reveal the following interesting characteristics: (1) an abnormal GC dinucleotide sequence located at the 5'-splicing site of the second intron of α D globins instead of a consensus GT-this finding is common to all studied Geochelone species; (2) a repeated sequence 5'-GCCCCGCGCCCCGC-3' found only in the first intron of the G. nigra α A globin gene, is a unique feature distinguishing the Galapagos giant tortoise from the other Geochelone tortoises that have non-repeated GCCCCGC sequences.

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Section: Molecular Relationships and Divergence Timessupporting

confidence: 89%

Section: Molecular Relationships and Divergence Timesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Globin-intron Sequences Imply Molecular Relationships Between the Galapagos Giant Tortoise and the Aldabra Giant Tortoise

Shishikura

2013

Nichidai Igaku Zasshi

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“…The Podocnemis red blood cells contain two kinds of hemoglobin components of reptilian species described in earlier studies (10,(17)(18)(19)(20). The major component, Hb A, separates easily from hemoglobin solution containing Hb D which is less abundant and tends to polymerize in preparation (21).…”

Section: Primary Structures Of a A -And B-globinsmentioning

confidence: 98%

“…Prior to separation of a-globin and b-globin, the purified Hb A was S-pyridylethylated by the method described previously (11) The modified Hb A was charged onto a reversed-phase column (Resource RPC, GE HealthCare) that was eluted with a buffered gradient of 0.1% trifluoroacetic acid (TFA) to 60% acetonitrile with 0.08% TFA. The other conditions were almost identical to those described previously (10): the fractions were monitored at 215 nm and 280 nm by spectrophotometer equipped with an Ä KTA purifier system (GE HealthCare). The N-terminal amino acid sequencing analysis of a A -and b-globins was performed using a gas phase protein sequencer, PPSQ-10 (Shimadzu, Shiga, Japan), equipped with a class LC-10 amino acid analyzer (Shimadzu).…”

Section: Primary Structure Of Hb Amentioning

confidence: 99%

Side‐necked turtle (Pleurodira, Chelonia, reptilia) hemoglobin: cDNA‐derived primary structures and X‐ray crystal structures of Hb A

2011

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Red blood cells of yellow-spotted river turtles (Podocnemis unifilis, Pleurodira, Chelonia, REPTILIA) have two hemoglobin (Hb) components, Hb A and Hb D. We purified the hemoglobin component homologous to amniote (reptiles, birds, and mammals) adult Hb A which comprises two identical α(A) -globin polypeptides and two identical β-globin polypeptides. To establish the crystal structure of Podocnemis Hb A, we first determined the globin primary structures using cDNA nucleotide sequencing with the assistance of protein sequencing. The purified Podocnemis Hb A produced a different form of crystal for each of the two different buffer systems used: form A, tetragonal crystals (space group, P4₁2₁2), produced under neutral pH (pH 7-8) conditions; and form B, hexagonal crystals (space group, P6₁22), produced under high alkaline pH (pH 11-13) conditions. Single crystals of the two forms were examined by Raman microscopy with an excitation of 532 nm, indicating their structural differences. The crystal structures of the two forms were constructed by X-ray crystallographic diffraction at a resolution of 2.20 Å for form A and 2.35 Å for form B. The differences of the tertiary and quaternary structures of the two forms were marginal; however, one clear difference was found in helix structure. When comparing Podocnemis Hb A with Hb A from specimens in other taxa, such as Anser indicus (birds) and Homo sapiens (mammals) by SHELXPRO, the root mean square deviation (RMSD) between the corresponding Cα atoms of the two globins does not exceed 2.0 Å. These low values indicate the crystal structures resemble each other. Our data on X-ray crystal structures and Raman spectra not only reveal the first findings on the two crystal forms of Podocnemis unifilis Hb A but also provide the first refined models for reptilian adult Hb A.

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Axolotl hemoglobin: cDNA-derived amino acid sequences of two α globins and a β globin from an adult Ambystoma mexicanum

Shishikura

Takeuchi

Nagai

2005

Comparative Biochemistry and Physiology Part B: Biochemistry an

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The Amino Acid Sequences of the α- and β-Globin Chains of Hemoglobin from the Aldabra Giant Tortoises, Geochelone gigantea

Cited by 8 publications

References 43 publications

Globin-intron Sequences Imply Molecular Relationships Between the Galapagos Giant Tortoise and the Aldabra Giant Tortoise

Globin-intron Sequences Imply Molecular Relationships Between the Galapagos Giant Tortoise and the Aldabra Giant Tortoise

Side‐necked turtle (Pleurodira, Chelonia, reptilia) hemoglobin: cDNA‐derived primary structures and X‐ray crystal structures of Hb A

Axolotl hemoglobin: cDNA-derived amino acid sequences of two α globins and a β globin from an adult Ambystoma mexicanum

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