An interaction between leukocyte cell-derived chemotaxin 2 and transferrin of ayu, Plecoglossus altivelis.
Yeast two-hybrid (Y2H) screens were used to test for interactions between leukocyte cell-derived chemotaxin 2 (LECT2) and a liver cDNA expression library of ayu, Plecoglossus altivelis. Of the 9 independent interacting clones identified, 7 were identical and closely related to transferrin (Tf) genes of fish, while the other two were related to c-type lectin genes. The interaction between ayu Tf (aTf) and ayu LECT2 (aLECT2) was confirmed by in vitro co-immunoprecipitation of the two proteins. Y2H assays using different parts of the two proteins showed that the segment aTf185-289 was not involved in the interaction with mature aLECT2, while the transit peptide of aLECT2 couldn't interact with entire aTf. Computer analysis revealed that aTf185-289, which contained two iron binding residues, Tyr197 and His253, was located at the N-terminus of aTf N-lobe. Strong interactions were also determined between LECT2 and Tf from the same animal, such as croceine croaker, Larimichthys crocea and mouse, Mus musculus. However, no cross-species interactions were determined. Based on published data, the Tf-LECT2 interaction is suggested to be most possibly involved in the body's defense against infection.
Chen
Jiong
J
Key Laboratory of the Ministry of Education for Applied Marine Biotechnology, Ningbo University, Ningbo City 315211, Zhejiang Province, People's Republic of China. jchen1975@163.com
Yang
Hong Y
HY
Shi
Yu H
YH
Li
Ming Y
MY
D000595:Amino Acid Sequence; D000818:Animals; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D007962:Leukocytes / Q000378:metabolism*; D008969:Molecular Sequence Data; D036441:Osmeriformes / Q000145:classification / Q000378:metabolism / Q000502:physiology*; D010802:Phylogeny; D011485:Protein Binding; D016415:Sequence Alignment; D014168:Transferrin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D036341:Intercellular Signaling Peptides and Proteins; D014168:Transferrin