Homologues of p48 Protein from Morula Cells of Ascidian Styela rustica in Other Species of Stolidobranchia

Tylets, M. I.; Daugavet, M. A.; Savelieva, A. V.; Podgornaya, O. I.; Shaposhnikova, T. G.

doi:10.1134/s1990519x19050080

Cited by 1 publication

(5 citation statements)

References 34 publications

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“…The composition of S. rustica cell fractions examined by phase-contrast microscopy was consistent with the results described previously 48 , 49 . In particular, fraction I enriched in hyaline amoebocytes, fraction II contained hyalinocytes and young morula cells, fraction III consisted of 95–97% of mature morula cells.…”

Section: Resultssupporting

confidence: 90%

“…In morula cells of S. rustica AB bound with 48 kDa and 26 kDa bands (Ref. 48 and present study Fig. 2 , I, II).…”

Section: Resultssupporting

confidence: 69%

“…SDS-PAGE was performed in 12% polyacrylamide gels with Unstained or Prestained Protein MW marker (Thermo Scientific Broad Range Unstained #26630, BioRad Prestained All Blue #1610393, Spectra™ Multicolor #26634) and stained with Coomassie BB G-250 (Biolot, Russia) or used to transfer proteins to a nitrocellulose membrane 0.2 μm or 0.45 µm (BioRad, #1620071, #1620117) for the Western blot 91 . Staining of S. rustica proteins was carried out as described previously 48 , 49 . For H. aurantium proteins the membrane was blocked with 3% solution of bovine serum albumin (BSA) in 1xPBS, 0.05% Tween for 1.5 h at RT and incubated with primary AB (GPαp48Sr 1:5000 in PBS-Tween) with the addition of 3% BSA overnight in + 10 °C.…”

Section: Methodsmentioning

confidence: 99%

“…During the repair process in the site of injury of ascidian S. rustica morula blood cells are the most abundant among all cell types 31 . It was shown that morula cells of S. rustica contain two cell-type specific proteins with molecular weights of 48 kDa (p48) and 26 kDa (p26) visible as major bands at SDS-electrophoresis of morula cells proteins 31 , 48 . Polyclonal antibodies (AB) against p48 interacted with both major proteins of morula cells demonstrating their alikeness.…”

Section: Introductionmentioning

confidence: 99%

“…AB also bind to the morula cells’ contents secreted into the tunic matrix 31 . Experiments on other ascidians from the order Stolidobranchia show that AB also bind to the morula cells and tunic matrix of Styela coriacea, Boltenia echinata, Molgula citrina and H. aurantium and test cells of reproductive system for all of them except H. aurantium 48 . In the present study, we described the gene, named Tuphoxin (Tunicate Phenol Oxidase), coding for p48 and several similar proteins in S. rustica and H. aurantium .…”

Section: Introductionmentioning

confidence: 99%

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New putative phenol oxidase in ascidian blood cells

Daugavet

Dobrynina

Shaposhnikova

et al. 2022

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The phenol oxidase system is ancient and ubiquitously distributed in all living organisms. In various groups it serves for the biosynthesis of pigments and neurotransmitters (dopamine), defence reactions and tissue hardening. Ascidians belong to subphylum Tunicata, which is considered the closest living relative to Vertebrates. Two phenol oxidases previously described for ascidians are vertebrate-like and arthropod-like phenol oxidases. In our present study, we described a new ascidian protein, Tuphoxin, with putative phenol oxidase function, which bears no sequence similarity with two enzymes described previously. The closest related proteins to Tuphoxin are mollusc haemocyanins. Unlike haemocyanins, which are oxygen transporting plasma proteins, Tuphoxin is synthesised in ascidian blood cells and secreted in the extracellular matrix of the tunic—ascidian outer coverings. Single mature transcript coding for this phenol oxidase can give several protein products of different sizes. Thus limited proteolysis of the initial protein is suggested. A unique feature of Tuphoxins and their homologues among Tunicata is the presence of thrombospondin first type repeats (TSP1) domain in their sequence which is supposed to provide interaction with extracellular matrix. The finding of TSP1 in the structure of phenol oxidases is new and we consider this to be an innovation of Tunicata evolutionary lineage.

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Section: Resultssupporting

confidence: 90%

“…In morula cells of S. rustica AB bound with 48 kDa and 26 kDa bands (Ref. 48 and present study Fig. 2 , I, II).…”

Section: Resultssupporting

confidence: 69%

Section: Methodsmentioning

confidence: 99%