TY - JOUR
T1 - Structural and functional characterization of Delphinus delphis hemoglobin system
AU - Manconi, Barbara
AU - Messana, Irene
AU - Maggiani, F
AU - Olianas, Alessandra
AU - Pellegrini, M
AU - Crnjar, R
AU - Castagnola, Massimo
AU - Giardina, Bruno
AU - Sanna, Mt
PY - 2009
Y1 - 2009
N2 - Structural analysis of the hemoglobin (Hb) system of Delphinus delphis revealed a high globin multiplicity: HPLC-electrospray ionization-mass spectrometry (ESI-MS) analysis evidenced three major beta (beta 1 16,022 Da, beta 2 16,036 Da, beta 3 16,036 Da, labeled according to their progressive elution times) and two major alpha globins (alpha 1 15,345 Da, alpha 2 15,329 Da). ESI-tandem mass and nucleotide sequence analyses showed that beta 2 globin differs from beta 1 for the substitution Val126 -> Leu, while beta 3 globin differs from beta 2 for the isobaric substitution Lys65 -> Gln. The alpha 2 globin differs from the alpha 1 for the substitution Ser15 -> Ala. Anion-exchange chromatography allowed the separation of two Hb fractions and HPLC-ESI-MS analysis revealed that the fraction with higher pI (HbI) contained beta 1, beta 2 and both the alpha globins, and the fraction with lower pI (HbII) contained beta 3 and both the alpha globins. Both D. delphis Hb fractions displayed a lower intrinsic oxygen affinity, a decreased effect of 2,3-BPG and a reduced cooperativity with respect to human HbA(0), with HbII showing the more pronounced differences. With respect to HbA(0), either the substitution Pro beta 5 -> Gly or the Pro beta 5 -> Ala is present in all the cetacean beta globins sequenced so far, and it has been hypothesized that position 5 of beta globins may have a role in the interaction with 2,3-BPG. Regarding the particularly lowered cooperativity of HbII, it is interesting to observe that the variant human HbA, characterized by the substitution Lys beta 65 -> Gln (HbJ-Cairo) has a decreased cooperativity with respect to HbA(0)
AB - Structural analysis of the hemoglobin (Hb) system of Delphinus delphis revealed a high globin multiplicity: HPLC-electrospray ionization-mass spectrometry (ESI-MS) analysis evidenced three major beta (beta 1 16,022 Da, beta 2 16,036 Da, beta 3 16,036 Da, labeled according to their progressive elution times) and two major alpha globins (alpha 1 15,345 Da, alpha 2 15,329 Da). ESI-tandem mass and nucleotide sequence analyses showed that beta 2 globin differs from beta 1 for the substitution Val126 -> Leu, while beta 3 globin differs from beta 2 for the isobaric substitution Lys65 -> Gln. The alpha 2 globin differs from the alpha 1 for the substitution Ser15 -> Ala. Anion-exchange chromatography allowed the separation of two Hb fractions and HPLC-ESI-MS analysis revealed that the fraction with higher pI (HbI) contained beta 1, beta 2 and both the alpha globins, and the fraction with lower pI (HbII) contained beta 3 and both the alpha globins. Both D. delphis Hb fractions displayed a lower intrinsic oxygen affinity, a decreased effect of 2,3-BPG and a reduced cooperativity with respect to human HbA(0), with HbII showing the more pronounced differences. With respect to HbA(0), either the substitution Pro beta 5 -> Gly or the Pro beta 5 -> Ala is present in all the cetacean beta globins sequenced so far, and it has been hypothesized that position 5 of beta globins may have a role in the interaction with 2,3-BPG. Regarding the particularly lowered cooperativity of HbII, it is interesting to observe that the variant human HbA, characterized by the substitution Lys beta 65 -> Gln (HbJ-Cairo) has a decreased cooperativity with respect to HbA(0)
KW - Amino acid sequence
KW - cDNA sequence
KW - Amino acid sequence
KW - cDNA sequence
UR - http://hdl.handle.net/10807/12352
U2 - 10.1007/s00360-009-0380-2
DO - 10.1007/s00360-009-0380-2
M3 - Article
SN - 0174-1578
VL - 179
SP - 971
EP - 983
JO - Journal of Comparative Physiology B: Biochemical, Systemic, and Environmental Physiology
JF - Journal of Comparative Physiology B: Biochemical, Systemic, and Environmental Physiology
ER -