A fatty acid-binding protein and a protein disulphide isomerase-related protein expressed in urochordate gonad cytosol

• Published in: The international journal of biochemistry & cell biology Vol. 32; no. 7; pp. 769 - 777
• Main Authors: Cavagnari, Brian M, Tatián, Marcos, Sahade, Ricardo J, Esnal, Graciela B, Santomé, José A
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.07.2000
• Subjects: Amino Acid Sequence; Animals; Cytosol - metabolism; Fatty acid-binding proteins; Fatty Acid-Binding Proteins - isolation & purification; Fatty Acid-Binding Proteins - metabolism; Gastrointestinal Tract - metabolism; Gonads - metabolism; Humans; Molecular Sequence Data; Molgula pedunculata; Protein Disulfide-Isomerases - isolation & purification; Protein Disulfide-Isomerases - metabolism; Protein disulphide isomerase; Sequence Alignment; Sequence comparison; Urochordata - metabolism; Urochordates; Sequence comparison; Fatty acid-binding proteins; Molgula pedunculata; Urochordates; Protein disulphide isomerase
• ISSN: 1357-2725; 1878-5875
• DOI: 10.1016/S1357-2725(00)00014-5

Despite the evolutionary-tree data suggesting that gene duplication leading to the divergence of the three branches which heart, liver and intestinal fatty acid-binding proteins belong to must have occurred before the vertebrate/invertebrate split, only the heart fatty acid-binding protein has been reported for invertebrates. In an attempt to shed light on this apparent inconsistency the presence of the other two branch members was investigated in the Urochordata Molgula pedunculata, an ascidian species close to vertebrates. The mantle-, gonad- and digestive tube-cytosolic fractions, obtained by centrifugation at 106,000  g , were incubated separately with [1- 14C]palmitic acid and then fractionated on a Sephadex G-75 column. In the case of gonads and digestive tube, radioactive peaks corresponding to a molecular mass of 14–16 kDa, characteristic of fatty acid-binding proteins, were detected. When the experiment was performed on the mantle, this peak showing fatty acid binding capacity was absent. Western Blot of the radioactive 14–16 kDa Sephadex fraction from the urochordate gonad cross-reacted with rat liver fatty acid-binding protein anti-serum but did not do so with anti-rat intestinal, adipocyte or heart fatty acid-binding protein antisera. The material from the digestive tube was not recognized by any of the antisera. The most abundant protein in said 14–16 kDa fraction was a protein disulphide isomerase-related protein. Its partial amino acid sequence was determined.