Purification, Partial Characterization and Peptide Sequences of Vitellogenin from a Reptile, the Tuatara (Sphenodon punctatus)

Brown, Mark; Carne, Alan; Chambers, Geoffrey K.

doi:10.1016/s0305-0491(96)00317-3

Cited by 17 publications

(9 citation statements)

References 55 publications

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“…The molecular mass of the VTG monomers from both species is consistent with structural information available on VTG from numerous fishes (Specker and Sullivan 1994) including carp (wide 150 kDa band, Folmar et al 1996; wide 156 kDa band, Fukada et al 2003;150 kDa, Hennies et al 2003;150 KDa, Nilsen et al 2004). Similar results were observed by SDS-PAGE analysis in non-denaturing conditions (data not shown), suggesting that the VTG monomers are not linked by interchain disulfide bonds in the native homodimeric form, as has been reported for other oviparous vertebrates (de Vlaming et al 1980;Brown et al 1997).…”

Section: Sds-pagesupporting

confidence: 88%

Purification and partial characterization of vitellogenin from shorthead redhorse (Moxostoma macrolepidotum) and copper redhorse (Moxostoma hubbsi) and detection in plasma and mucus with a heterologous antibody

Maltais

2008

Fish Physiol Biochem

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Vitellogenin (VTG), the egg yolk precursor protein, was purified from plasma of estradiol-3-benzoate (E2B)-treated male shorthead redhorse (Moxostoma macrolepidotum) and immature copper redhorse (Moxostoma hubbsi) by a two-step chromatographic procedure without precipitation. Intact VTGs appeared as dimers with apparent molecular masses, determined by gel filtration, of approximately 425 kDa (copper redhorse) and approximately 450 kDa (shorthead redhorse). In native polyacrylamide gel electrophoresis (PAGE), dimeric redhorse VTGs appeared as a 520 kDa band. Both VTGs were reduced to a single monomer of approximately 150 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing and nonreducing conditions, indicating that monomers are not linked by disulfide bonds in the dimer form. The purified proteins were characterized as phospholipoglycoproteins. Isoelectric focusing of both VTGs revealed components with isoelectric points ranging from 5.3 to 6.0, suggesting charge heterogeneity. The amino acid composition of both VTGs contains a high proportion of nonpolar amino acids and was similar to those of other teleosts. An antibody developed against carp (Cyprinus carpio) VTG showed cross-reactivity with VTG from both redhorse species. Using this antibody, VTG was detected in plasma and surface mucus of E2B-treated redhorse. This is the most extensive report on purification and characterization of vitellogenin from catostomidid species.

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Section: Sds-pagesupporting

confidence: 88%

Purification and partial characterization of vitellogenin from shorthead redhorse (Moxostoma macrolepidotum) and copper redhorse (Moxostoma hubbsi) and detection in plasma and mucus with a heterologous antibody

Maltais

2008

Fish Physiol Biochem

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“…The predicted protein sequence contains all the conserved domains of the VTG family (Figure ), indicating the full‐length vtg cDNA sequence of S. formosus belongs to the vitellogenin gene family. The encoded protein sequence contained a conserved vitellogenin‐N domain, consistent with the results of Brown, Carne, and Chambers () who found that the N‐terminal sequence of vitellogenin was highly conserved among chordates. Further, the sequence analysis showed that the vitellogenin‐N domain was an area of lipoprotein, which contains two structures of the LV1n chain.…”

Section: Discussionsupporting

confidence: 88%

Tissue expression and bioinformatics analysis of the vitellogenin gene of Asian arowana ( Scleropages formosus )

et al. 2019

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The RACE technique was used to clone the full‐length vitellogenin (VTG) cDNA sequence of Asian arowana (Scleropages formosus). The full‐length sequence was 5,550 bp with an open reading frame of 5,238 bp, encoding 1,745 amino acids, and 5′ and 3′ UTRs (untranslated regions) of 45 bp and 267 bp, respectively. Phylogenetic analysis showed that S. formosus and silver arowana (Osteoglossum bicirrhosum) share a close evolutionary relationship (bootstrap 100%). The quantitative real‐time PCR results showed that vtg expression was significantly higher in liver and gonads of male and female fish compared with its expression in the other tissues tested (p < 0.01). The relative expression levels of vtg in liver, gland, kidney, heart, head kidney, and brain of female fish were significantly higher than in the corresponding tissues of male fish (p < 0.05).

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“…Sequence alignment was achieved using the BLAST network service. No registers of any reptile VTG sequence were found in the NCBI database, but Herbst et al [20] and Brown et al [31] previously reported the N‐terminal sequence of VTG from the green turtle ( Chelonia mydas ) and the tuatara ( Sphen‐odon punctatus ), respectively. In C. mydas agassizii , the N‐terminal sequence alignment of the 200‐kDa protein was identical to the VTG sequence from C. mydas and similar in eight amino acids to the VTG sequence from S. punctatus (Table 2).…”

Section: Resultsmentioning

confidence: 99%

“…Similarly, when the E 2 ‐treated plasma was run on a DEAE‐Sepharose column along with a linear salt gradient, it was shown by native‐PAGE that the inducible peak contained only the 240‐ and 500‐kDa IPs. Moreover, the two IPs eluted at a high salt concentration (360 mM), providing evidence of their high phosphorous composition, a consistent characteristic of reptilian VTG [18,31,32]. Degradation of the 500‐kDa protein was observed when it was injected onto the column in the absence of PMSF, resulting in the presence of the 240‐kDa protein.…”

Section: Discussionmentioning

confidence: 99%

Vitellogenin in black turtle (Chelonia mydas agassizii): Purification, partial characterization, and validation of an enzyme-linked immunosorbent assay for its detection

Sifuentes-Romero

Vázquez-Boucard

Sierra-Beltrán

et al. 2006

Environmental Toxicology and Chemistry

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Black turtle plasmatic vitellogenin (VTG) was purified from 17beta-estradiol-induced males using ion-exchange chromatography. The isolated protein was identified as VTG by its glycolipoprotein nature and amino acid sequence homology with other vertebrate VTG. It was characterized as a 500-kDa dimer composed of two identical, 200- to 240-kDa monomers. Polyclonal antibodies raised against black turtle VTG showed high titer and specificity, as demonstrated by enzyme-linked immunosorbent assay and Western blot analysis, respectively. The range of the assay was estimated to be between 15 ng/ml and 2 microg/ml, and the inter- and intra-assay coefficients of variation were 9.4 and 7.3%, respectively. Black turtle antibody cross-reacted with VTG of two other sea turtle species, Caretta caretta (loggerhead) and Eretmochelys imbricata (hawksbill), extending the applicability of the assay as part of a sea turtle health assessment program.

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Purification, Partial Characterization and Peptide Sequences of Vitellogenin from a Reptile, the Tuatara (Sphenodon punctatus)

Cited by 17 publications

References 55 publications

Purification and partial characterization of vitellogenin from shorthead redhorse (Moxostoma macrolepidotum) and copper redhorse (Moxostoma hubbsi) and detection in plasma and mucus with a heterologous antibody

Purification and partial characterization of vitellogenin from shorthead redhorse (Moxostoma macrolepidotum) and copper redhorse (Moxostoma hubbsi) and detection in plasma and mucus with a heterologous antibody

Tissue expression and bioinformatics analysis of the vitellogenin gene of Asian arowana ( Scleropages formosus )

Vitellogenin in black turtle (Chelonia mydas agassizii): Purification, partial characterization, and validation of an enzyme-linked immunosorbent assay for its detection

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