Immunohistochemical studies of human ribosomal protein S3 (rpS3).
The human ribosomal protein S3 (rpS3) was expressed in E. coli using the pET-15b vector and the monoclonal antibodies (mAbs) were produced and characterized. A total of five hybridoma cell lines were established and the antibodies recognized a single band of molecular weight of 33 kDa on immunoblot with purified rpS3. When the purified rpS3 was incubated with the mAbs, the UV endonuclease activity of rpS3 was inhibited up to a maximum of 49%. The binding affinity of mAbs to rpS3 determined by using a biosensor technology showed that they have similar binding affinities. Using the anti-rpS3 antibodies as probes, we investigated the cross-reactivities of various other mammalian brain tissues and cell lines, including human. The immunoreactive bands on Western blots appeared to be the same molecular mass of 33 kDa in all animal species tested. They also appear to be extensively cross-reactive among different organs in rat. These results demonstrated that only one type of immunologically similar rpS3 protein is present in all of the mammalian brain tissues including human. Furthermore, these antibodies were successfully applied in immunohistochemistry in order to detect rpS3 in the gerbil brain tissues. Among the various regions in the brain tissues, the rpS3 positive neurons were predominantly observed in the ependymal cells, hippocampus and stantia nigra pars compacta. The different distributions of rpS3 in brain tissues reply that rpS3 protein may play an important second function in the neuronal cells.
Department of Biomedical Sciences and Research Institute for Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Korea.
Soo Hyun
SH
Choi
So Young
SY
Kim
Jae Jin
JJ
An
Sun Hwa
SH
Lee
Dae Won
DW
Kim
Moo Ho
MH
Won
Tae-Cheon
TC
Kang
Jinseu
J
Park
Won Sik
WS
Eum
Joon
J
Kim
Soo Young
SY
Choi
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology / Q000494:pharmacology; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D045744:Cell Line, Tumor; D002645:Chickens; D004720:Endonucleases / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000528:radiation effects; D018604:Epitope Mapping; D005786:Gene Expression Regulation / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D013045:Species Specificity; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C044152:ribosomal protein S3; D004720:Endonucleases