Molecular cloning and characterization of tiger shrimp (Penaeus monodon) transglutaminase.
Transglutaminases (TG) are important for blood coagulation and post-translation remodeling of proteins. Using a plaque screening assay, we isolated cDNA encoding a novel TG from a shrimp (Penaeus monodon) hemocyte cDNA library. The TG cDNA consists of 2988 bp with an open reading frame of 2271 bp. The deduced protein has 757 amino acid residues, a calculated molecular mass of 84,713 Da and an isoelectric point of 5.56. Neither a typical hydrophobic leader sequence nor a transmembrane domain could be identified from the deduced sequence. Thus, shrimp TG may be a typical cytoplasmic protein. The sequence of shrimp TG was similar to crayfish, other invertebrate and vertebrate TG sequences. Enzyme activity was detected in all organs tested. This is consistent with the widespread, low-level expression of TG mRNA. However, high levels of TG expression were detected in hematopoietic tissue. TG signals were stronger in mitotic cells, indicating that cell proliferation and TG synthesis are associated. Preliminary data showed that recombinant TG existed the enzyme activity but lacked coagulation activity.
Huang
Chih-Cheng
CC
Institute of Zoology, National Taiwan University, Taipei 106, Taiwan, ROC.
Sritunyalucksana
Kallaya
K
Söderhäll
Kenneth
K
Song
Yen-Ling
YL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006434:Hemocytes / Q000201:enzymology; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000201:enzymology* / Q000235:genetics*; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D011503:Transglutaminases / Q000737:chemistry / Q000235:genetics*
D018076:DNA, Complementary; D012333:RNA, Messenger; D011503:Transglutaminases