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Characterization of a novel cellular retinoic acid/retinol binding protein from shrimp: expression of the recombinant protein for immunohistochemical detection and binding assay
- Source :
- Gene. 288:77-84
- Publication Year :
- 2002
- Publisher :
- Elsevier BV, 2002.
-
Abstract
- Members of the cellular retinoic acid (CRABP) and retinol binding (CRBP) proteins family are involved in the metabolic pathways of retinoic acid (RA) and retinal respectively. The objective of this study is to determine whether such proteins are present in crustaceans. We report here the cloning and isolation of a novel complementary DNA (cDNA) that showed characteristics of the CRABP/CRBP from the ovary and eyestalk of the shrimp. The cDNA is 0.9 Kb in size and the deduced shrimp protein is encoded for a protein of 14 kDa. Although it shows high amino acids sequence similarity to both the vertebrate and invertebrate CRABP, some conserved amino acids identified in other CRABPs were not found in MeCRABP. MeCRABP is expressed in the ovary, eyestalk, testis, epidermis and early larvae. The presence of MeCRABP in early larval stages suggests that the protein may be involved in the early larval development. Recombinant MeCRABP was produced and used to generate a polyclonal antibody. In the immunohistochemical detection study, anti-rCRABP antibody recognized the presence of CRABP in several cell types of the eyestalk as well as the smaller oocytes of the ovary. Although MeCRABP messenger RNA transcripts can be detected in the ovary throughout the ovarian maturation period, CRABP was detected only in the primary oocytes of the ovary. The results suggest that CRABP transcripts in the mature ovary are not translated and may be supplied to the oocyte as maternal messages. The binding property of the recombinant MeCRABP was also tested by a fluorometeric method. The result indicates that rMeCRABP binds to both RA and retinal with similar affinity. This study represents the first cloning and characterization of a cDNA that belongs to a member of retinoid/fatty acid binding protein family in crustaceans.
- Subjects :
- DNA, Complementary
Molecular Sequence Data
Retinoic acid
Gene Expression
Tretinoin
Biology
Binding, Competitive
Antibodies
Fatty acid-binding protein
chemistry.chemical_compound
Decapoda
Complementary DNA
Gene expression
Genetics
Animals
Amino Acid Sequence
Cloning, Molecular
Vitamin A
Peptide sequence
Phylogeny
Messenger RNA
Base Sequence
Sequence Homology, Amino Acid
Retinol-Binding Proteins, Cellular
Sequence Analysis, DNA
General Medicine
Immunohistochemistry
Molecular biology
Recombinant Proteins
Retinol-Binding Proteins
Retinol binding protein
chemistry
Biochemistry
Rabbits
Retinol binding
Sequence Alignment
Subjects
Details
- ISSN :
- 03781119
- Volume :
- 288
- Database :
- OpenAIRE
- Journal :
- Gene
- Accession number :
- edsair.doi.dedup.....44c125604ceaf01ba32e85e0f745511a