Fine Sampling of the R→T Quaternary‐Structure Transition of a Tetrameric Hemoglobin

Vitagliano, Luigi; Mazzarella, L.; Merlino, Antonello; Vergara, Alessandro

doi:10.1002/chem.201603421

Cited by 9 publications

(19 citation statements)

References 68 publications

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“…4) and has never been detected in the structures of other globins such as Mb, Cygb and Ngb (data not shown). Although the overall architecture of the distal side of the β-heme pocket is essentially preserved, it is worth mentioning that, as occasionally observed in other Hbs 11,39 , the distal His63β swings out of the heme pocket (Fig. S1B).…”

Section: Resultssupporting

confidence: 60%

“…In this context, the characterization of Hbs isolated from organisms living in extreme conditions has proven to be particularly fruitful. Indeed, studies carried out on Hbs of fish thriving in the freezing water of the Antarctic Ocean have provided interesting insights into the repertoire of structural states that Hbs may assume as well as into the variety of oxidation states that can characterize the heme iron 11–28 . The Southern Ocean, which surrounds Antarctica, is the coldest ocean on Earth and it is isolated from the other oceans by the Antarctic circumpolar current in the last ∼32 million years 29 .…”

Section: Introductionmentioning

confidence: 99%

“…Its genome, recently sequenced, is the best representative of the temperate character of the most recent common ancestor of the Antarctic notothenioids 33 . This species is a close relative of the Antarctic notothenioids such as Trematomus bernacchii 12,15,16,23,28,34 and Trematomus newnesi 11,13,19,35 whose Hbs have been extensively characterized. The spectroscopic characterization of E. maclovinus Hb1 has unraveled that the protein presents two distinct CO forms in solution 36,37 .…”

Section: Introductionmentioning

confidence: 99%

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The unique structural features of carbonmonoxy hemoglobin from the sub-Antarctic fish Eleginops maclovinus

Balasco¹,

Vitagliano²,

Merlino³

et al. 2019

Sci Rep

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Tetrameric hemoglobins (Hbs) are prototypical systems for the investigations of fundamental properties of proteins. Although the structure of these proteins has been known for nearly sixty years, there are many aspects related to their function/structure that are still obscure. Here, we report the crystal structure of a carbonmonoxy form of the Hb isolated from the sub-Antarctic notothenioid fish Eleginops maclovinus characterised by either rare or unique features. In particular, the distal site of the α chain results to be very unusual since the distal His is displaced from its canonical position. This displacement is coupled with a shortening of the highly conserved E helix and the formation of novel interactions at tertiary structure level. Interestingly, the quaternary structure is closer to the T-deoxy state of Hbs than to the R-state despite the full coordination of all chains. Notably, these peculiar structural features provide a rationale for some spectroscopic properties exhibited by the protein in solution. Finally, this unexpected structural plasticity of the heme distal side has been associated with specific sequence signatures of various Hbs.

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Section: Resultssupporting

confidence: 60%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The unique structural features of carbonmonoxy hemoglobin from the sub-Antarctic fish Eleginops maclovinus

Balasco¹,

Vitagliano²,

Merlino³

et al. 2019

Sci Rep

Self Cite

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“…In the deoxygenated form, the distal histidine side chains have a propensity to swing out of the heme pocket, thus allowing a compression of the surrounding helices with respect to each other, which in turn causes Fe 2+ to move out of the porphyrin plane [64]. This gives rise to a tensed conformation ("T" conformation) with the heme adopting a dome-like arrangement ( Figure 2).…”

Section: Hemoglobins: Structure Function and Aggregationmentioning

confidence: 99%

Rational Drug Design of Peptide-Based Therapies for Sickle Cell Disease

2019

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Sickle cell disease (SCD) is a group of inherited disorders affecting red blood cells, which is caused by a single mutation that results in substitution of the amino acid valine for glutamic acid in the sixth position of the β-globin chain of hemoglobin. These mutant hemoglobin molecules, called hemoglobin S, can polymerize upon deoxygenation, causing erythrocytes to adopt a sickled form and to suffer hemolysis and vaso-occlusion. Until recently, only two drug therapies for SCD, which do not even fully address the manifestations of SCD, were approved by the United States (US) Food and Drug Administration. A third treatment was newly approved, while a monoclonal antibody preventing vaso-occlusive crises is also now available. The complex nature of SCD manifestations provides multiple critical points where drug discovery efforts can be and have been directed. These notwithstanding, the need for new therapeutic approaches remains high and one of the recent efforts includes developments aimed at inhibiting the polymerization of hemoglobin S. This review focuses on anti-sickling approaches using peptide-based inhibitors, ranging from individual amino acid dipeptides investigated 30-40 years ago up to more promising 12-and 15-mers under consideration in recent years.Molecules 2019, 24, 4551 2 of 23 with hemoglobin reduces HbS solubility and promotes polymerization, also called sickling [3,4]. This ultimately leads to hampered O 2 binding and transport, impaired erythrocyte morphology and interaction with endothelial surfaces [5,6], premature erythrocyte rupture and anemia, painful vaso-occlusive crisis, a general poor health, and, in many cases, death [7][8][9][10][11].Despite growing understanding of the polymerization of HbS and its effects on red blood cells (RBCs), until very recently, only two drugs-hydroxyurea and L-glutamine-were approved by the United States (US) Food and Drug Administration (FDA) for the management of SCD [12]. Hydroxyurea is the most widely employed drug treatment of sickle cell anemia in different age groups [13][14][15][16][17][18]. While its clinically observed efficacy has been attributed to different effects at the cellular level [19], the most important mechanism of action relates to its ability to induce the production of fetal hemoglobin (HbF), which does not polymerize, and to increase the total concentration of hemoglobin [20,21]. Hydroxyurea remains a viable treatment option for SCD, and the concern of toxicities associated with its administration has largely been limited to side effects that resolve with medication discontinuation [22][23][24][25][26]. There have, however, been certain reports of associated malignancies [27][28][29][30][31][32], but further investigations are needed to categorically confirm these [33]. L-glutamine is the second approved drug treatment [12,34]. While its mechanism of action is not known, and only suggested to involve a reduction of oxidative stress via elevation of the levels of reduced glutathione [35,36], it is clear that it has no effect on hemo...

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“…In this manuscript, we extend our hypothesis and report that: (i) ELP phase separation can also detect large (60 kDa) tetrameric proteins; and (ii) this can be tuned to occur at physiological temperatures even with a high molecular weight ELP (77 kDa) (Table ). Due to the ubiquity of multimeric proteins in nature, including IL‐5 cytokine (dimer), hemoglobin (tetramer), immunoglobulins of class IgA (dimer) and IgM (pentamer), and cell‐surface receptor tyrosine kinases (dimer) like VEGFR and EGFR, crosslinking ELPs might be designed to detect many species important in diagnostic and therapeutic applications.…”

Section: Introductionmentioning

confidence: 99%

Elastin‐like polypeptide switches: A design strategy to detect multimeric proteins

et al. 2017

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Elastin-Like Polypeptides (ELPs) reversibly phase separate in response to changes in temperature, pressure, concentration, pH, and ionic species. While powerful triggers, biological microenvironments present a multitude of more specific biological cues, such as antibodies, cytokines, and cell-surface receptors. To develop better biosensors and bioresponsive drug carriers, rational strategies are required to sense and respond to these target proteins. We recently reported that noncovalent association of two ELP fusion proteins to a "chemical inducer of dimerization" small molecule (1.5 kDa) induces phase separation at physiological temperatures. Having detected a small molecule, here we present the first evidence that ELP multimerization can also detect a much larger (60 kDa) protein target. To demonstrate this strategy, ELPs were biotinylated at their amino terminus and mixed with tetrameric streptavidin. At a stoichiometric ratio of [4:1], two to three biotin-ELPs associate with streptavidin into multimeric complexes with an apparent K of 5 nM. The increased ELP density around a streptavidin core strongly promotes isothermal phase separation, which was tuned to occur at physiological temperature. This phase separation reverses upon saturation with excess streptavidin, which only favors [1:1] complexes. Together, these findings suggest that ELP association with multimeric biomolecules is a viable strategy to deliberately engineer ELPs that respond to multimeric protein substrates.

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Fine Sampling of the R→T Quaternary‐Structure Transition of a Tetrameric Hemoglobin

Cited by 9 publications

References 68 publications

The unique structural features of carbonmonoxy hemoglobin from the sub-Antarctic fish Eleginops maclovinus

The unique structural features of carbonmonoxy hemoglobin from the sub-Antarctic fish Eleginops maclovinus

Rational Drug Design of Peptide-Based Therapies for Sickle Cell Disease

Elastin‐like polypeptide switches: A design strategy to detect multimeric proteins

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