A Heme-binding Protein from Hemolymph and Oocytes of the Blood-sucking Insect, Rhodnius prolixus

Oliveira, Pedro L.; Kawooya, John K.; Ribeiro, José M. C.; Meyer, Terrance E.; Poorman, Roger A.; Alves, Elias W.; Walker, F. Ann; Machado, Ednildo A.; Nussenzveig, Roberto; Padovan, Gilberto João; Masuda, Hatisaburo

doi:10.1074/jbc.270.18.10897

Cited by 88 publications

(64 citation statements)

References 22 publications

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“…For example, the iron transport protein transferrin ($65 kDa) is selectively deposited in the yolk of the flesh fly, Sarcophaga peregrina (Kurama et al, 1995), and a similar protein is deposited in the bean bug Riptortus clavatus (Hirai et al, 2000). The hemolymph and oocytes of R. prolixus also contain a 15 kDa heme-binding protein (Oliveira et al, 1995). In Lepidoptera, a blue biliprotein called insecticyanin, a component of larval hemolymph, is deposited in developing oocytes and is present in mature eggs (Chino et al, 1969;Kang et al, 1995).…”

Section: Ovarian Yolk Proteinsmentioning

confidence: 99%

Vitellogenesis and Post-Vitellogenic Maturation of the Insect Ovarian Follicle

Swevers

2005

Comprehensive Molecular Insect Science

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Section: Ovarian Yolk Proteinsmentioning

confidence: 99%

Vitellogenesis and Post-Vitellogenic Maturation of the Insect Ovarian Follicle

Swevers

2005

Comprehensive Molecular Insect Science

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“…However, when Pd-mP was bound to the globin polypeptide, its red emission at 665·nm greatly increased in comparison to Pd-mP in methanol or water (Fig.·3B). Several other proteins were tested, including bovine albumin, HeLP, a heme-binding lipoprotein from the Boophilus microplus hemolymph (MayaMonteiro et al, 2000) and RHBP, a heme-binding protein from Rhodnius prolixus (Oliveira et al, 1995), and all of them induced a fluorescence peak at 665·nm, while proteins not capable of binding heme, such as bovine trypsin, did not alter the emission profile (not shown). Phospholipid liposomes, on the other hand, showed an intermediate profile, with a 665·nm fluorescence peak higher than that obtained in water or methanol, but much less intense than that of protein-bound PdmP (Fig.·3B).…”

Section: Palladium Mesoporphyrin IX (Pd-mp) Fluorescencementioning

confidence: 99%

Tracing heme in a living cell: hemoglobin degradation and heme traffic in digest cells of the cattle tickBoophilus microplus

Lara

Lins

Bechara

et al. 2005

Journal of Experimental Biology

128

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Heme is present in all cells, acting as a cofactor in essential metabolic pathways such as respiration and photosynthesis. Moreover, both heme and its degradation products, CO, iron and biliverdin, have been ascribed important signaling roles. However, limited knowledge is available on the intracellular pathways involved in the flux of heme between different cell compartments. The cattle tick Boophilus microplus ingests 100 times its own mass in blood. The digest cells of the midgut endocytose blood components and huge amounts of heme are released during hemoglobin digestion. Most of this heme is detoxified by accumulation into a specialized organelle, the hemosome.We followed the fate of hemoglobin and albumin in primary cultures of digest cells by incubation with hemoglobin and albumin labeled with rhodamine. Uptake of hemoglobin by digest cells was inhibited by unlabeled globin, suggesting the presence of receptor-mediated endocytosis. After endocytosis, hemoglobin was observed inside large digestive vesicles. Albumin was exclusively associated with a population of small acidic vesicles, and an excess of unlabeled albumin did not inhibit its uptake. The intracellular pathway of the heme moiety of hemoglobin was specifically monitored using Palladium-mesoporphyrin IX (Pd-mP) as a fluorescent heme analog. When pulse and chase experiments were performed using digest cells incubated with Pd-mP bound to globin (Pd-mP-globin), strong yellow fluorescence was found in large digestive vesicles 4·h after the pulse. By 8·h, the emission of Pd-mP was red-shifted and more evident in the cytoplasm, and at 12·h most of the fluorescence was concentrated inside the hemosomes and had turned green. After 48·h, the Pd-mP signal was exclusively found in hemosomes. In methanol, Pd-mP showed maximal emission at 550·nm, exhibiting a red-shift to 665·nm when bound to proteins in vitro.The red emission in the cytosol and at the boundary of hemosomes suggests the presence of heme-binding proteins, probably involved in transport of heme to the hemosome. The existence of an intracellular heme shuttle from the digestive vesicle to the hemosome acting as a detoxification mechanism should be regarded as a major adaptation of ticks to a blood-feeding way of life. To our knowledge, this is the first direct observation of intracellular transport of heme in a living eukaryotic cell. A similar approach, using Pd-mP fluorescence, could be applied to study heme intracellular metabolism in other cell types.

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“…Binding Studies-Binding of heme to THAP was monitored by measuring light absorption at 411 nm while progressively adding a solution of 0.1 mM hemin in 0.1 M NaOH (24). The absorbance was plotted against the amount of hemin added, and the hemin necessary to fully saturate the heme binding capacity of the protein was determined from the break in the curve.…”

Section: Fig 3 Amino Acid Sequence Of Thapmentioning

confidence: 99%

A Heme-binding Aspartic Proteinase from the Eggs of the Hard TickBoophilus microplus

Sorgine

Logullo

Zingali

et al. 2000

Journal of Biological Chemistry

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An aspartic proteinase that binds heme with a 1:1 stoichiometry was isolated and cloned from the eggs of the cattle tick Boophilus microplus. This proteinase, herein named THAP (tick heme-binding aspartic proteinase) showed pepstatin-sensitive hydrolytic activity against several peptide and protein substrates. Although hemoglobin was a good substrate for THAP, low proteolytic activity was observed against globin devoid of the heme prosthetic group. Hydrolysis of globin by THAP increased as increasing amounts of heme were added to globin, with maximum activation at a heme-toglobin 1:1 ratio. Further additions of heme to the reaction medium inhibited proteolysis, back to a level similar to that observed against globin alone. The addition of heme did not change THAP activity toward a synthetic peptide or against ribonuclease, a non-hemeprotein substrate. The major storage protein of tick eggs, vitellin (VT), the probable physiological substrate of THAP, is a hemeprotein. Hydrolysis of VT by THAP was also inhibited by the addition of heme to the incubation media. Taken together, our results suggest that THAP uses heme bound to VT as a docking site to increase specificity and regulate VT degradation according to heme availability.

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A Heme-binding Protein from Hemolymph and Oocytes of the Blood-sucking Insect, Rhodnius prolixus

Cited by 88 publications

References 22 publications

Vitellogenesis and Post-Vitellogenic Maturation of the Insect Ovarian Follicle

Vitellogenesis and Post-Vitellogenic Maturation of the Insect Ovarian Follicle

Tracing heme in a living cell: hemoglobin degradation and heme traffic in digest cells of the cattle tickBoophilus microplus

A Heme-binding Aspartic Proteinase from the Eggs of the Hard TickBoophilus microplus

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