Determination of phosphorylated amino acid residues of Rab8 from <i>Bombyx mori</i>

Uno, Tomohide; Nakada, Takuya; Okamaoto, Sota; Nakamura, Masahiko; Matsubara, Mamoru; Imaishi, Hiromasa; Yamagata, Hiroshi; Kanamaru, Kengo; Takagi, Michihiro

doi:10.1002/arch.20201

Cited by 13 publications

(10 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Ser-132, is also phosphorylated by PKC in vitro (Uno et al, 2007). The present study showed that BRab1, BRab7, BRab10 and BRab11, in addition to BRab8, were phosphorylated by PKC in vitro (Fig.…”

Section: Discussionsupporting

confidence: 64%

See 1 more Smart Citation

Phosphorylation of small GTPase Rab proteins from Bombyx mori (Lepidoptera: Bombycidae)

Uno¹,

Hata²,

Thi³

et al. 2009

Eur. J. Entomol.

Self Cite

View full text Add to dashboard Cite

Abstract. Small GTPases of the Rab family act as essential regulators of vesicle transport pathways. Five Rab cDNA clones (BRab1, 7, 8, 11 and 14) from Bombyx mori were expressed in Escherichia coli as a thioredxin or glutathione sulfotransferase fusion protein. After purification, the fusion protein was tested for phosphorylation using protein kinase C (PKC). Results indicate that all of them were phosphorylated in vitro. The phosphorylation site of BRab1 was determined by mass-spectrometric analysis, which identified that Ser-17 of BRab1 was phosphorylated by PKC. Deletion and site-directed mutagenesis indicated that Ser-111of BRab8, in addition to Ser-17, was newly phosphorylated. Further immunohistochemical analysis using antibodies against Rab8 indicated that there are some Rab8 immunoreactive cells close to the neuropeptide secreting cells. This result suggests that in insects Rab proteins are regulated by phosphorylation and at least some of them are involved in neuropeptide secretion.

show abstract

Section: Discussionsupporting

confidence: 64%

“…Ser-132 of BRab8 is phosphorylated by PKC (Uno et al, 2007). However, phosphorylatd amino acid residues of BRab8 were not identified by the MS analysis (data not shown).…”

Section: Phosphorylation Of Fragments Of Brab8mentioning

confidence: 92%

Phosphorylation of small GTPase Rab proteins from Bombyx mori (Lepidoptera: Bombycidae)

Uno¹,

Hata²,

Thi³

et al. 2009

Eur. J. Entomol.

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Introductionmentioning

confidence: 99%

“…Only Uno et al (2006Uno et al ( , 2007 have shown that Rab8 from Bombyx mori (BRab8) is phosphorylated in vitro by recombinant B. mori PKC and by protein kinases purified from the brain of B. mori. They have determined one of the phosphorylation sites of BRab8 as being Ser-132 (Uno et al 2007). In order to examine the possible regulation of neurosecretion via the phosphorylation of Rab8 by PKC, the present investigation has targeted Rab8, prothoracicotropic hormone (PTTH), and PKC in the brain to determine the functional roles of this protein in neuropeptide secretion in the American cockroach Periplaneta americana.…”

Section: Introductionmentioning

confidence: 99%

Putative regulatory mechanism of prothoracicotropic hormone (PTTH) secretion in the American cockroach, Periplaneta americana as inferred from co-localization of Rab8, PTTH, and protein kinase C in neurosecretory cells

2009

Self Cite

View full text Add to dashboard Cite

Small GTPases of the Rab family act as essential regulators of vesicle transport pathways, including the exocytosis of neurohormones. These processes are not well-understood in insects. To address the physiological function of Rab proteins and their phosphorylation in insect neurosecretion, Rab8-like, prothoracicotropic hormone (PTTH)-like, and protein kinase C (PKC)-like immunohistochemical reactivities (-ir) were investigated in the brain of the American cockroach, Periplaneta americana. All the antibodies tested reacted with neurons in the pars intercerebralis, corpora cardiaca, and nervi corporis allati I. Double-labeling experiments demonstrated that all PTTH-ir were colocalized with Rab8-ir and PKC-ir in the pars intercerebralis, although exclusive reactivity was present to antisera against Rab8 or PKC. These findings support the notion that Rab8-like antigen is phosphorylated by PKC, and that this phosphorylation is involved in the axonal transport and secretion of PTTH in this species.

show abstract

“…Only a series of experiment suggests a function of Rab protein insect brain. Bombyx mori (BRab8) was phosphorylated by B. mori protein kinase C (PKC) (Uno et al 2006, Uno et al 2007, followed up with the co-localization experiment of Rab8 with PKC in insect neurosecretory cells (Hiragaki et al 2009). …”

Section: Introductionmentioning

confidence: 99%

Small GTPases of the Rab family in the brain of Bombyx mori

Uno

Hata

Hiragaki

et al. 2010

Histochem Cell Biol

Self Cite

View full text Add to dashboard Cite

Small GTPases of the Rab family are key regulators of membrane trafficking. We produced antibodies against the Rab7 protein of Bombyx mori (BRab7) in rabbits, and against the Rab11 protein of B. mori (BRab11) in mice. The antibodies recognized BRab7 and BRab11 proteins, but did not recognize other Rab proteins. Immunoblotting of samples from brain tissue of B. mori revealed a single band for each antibody. Rab11 was expressed in most tissues, whereas Rab7 was expressed in the brain, ovary, and testis. Immunohistochemical reactivity of Rab7 and Rab11 in the brain of B. mori was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Double-labeling experiments demonstrated that immunohistochemical reactivity of Rab7 co-localized with that of Rab11 and partially with that of Rab8. Immunohistochemical reactivity of Rab11 and Rab8 co-localized with that of PERIOD, one of the proteins associated with circadian rhythm. These findings suggest that Rab7, Rab8, and Rab11 are involved in protein transport in the neurons of the brain of B. mori and might play a role in the control of circadian rhythm.

show abstract

Determination of phosphorylated amino acid residues of Rab8 from Bombyx mori

Cited by 13 publications

References 42 publications

Phosphorylation of small GTPase Rab proteins from Bombyx mori (Lepidoptera: Bombycidae)

Phosphorylation of small GTPase Rab proteins from Bombyx mori (Lepidoptera: Bombycidae)

Putative regulatory mechanism of prothoracicotropic hormone (PTTH) secretion in the American cockroach, Periplaneta americana as inferred from co-localization of Rab8, PTTH, and protein kinase C in neurosecretory cells

Small GTPases of the Rab family in the brain of Bombyx mori

Contact Info

Product

Resources

About