title: Subcellular dissemination of prothymosin alpha at normal physiology: immunohistochemical vis-a-vis western blotting perspective.
abstract: BACKGROUND
  The cell type, cell status and specific localization of Prothymosin α (PTMA) within cells seemingly determine its function. PTMA undergoes 2 types of protease proteolytic modifications that are useful in elucidating its interactions with other molecules; a factor that typifies its roles. Preferably a nuclear protein, PTMA has been shown to function in the cytoplasm and extracellularly with much evidence leaning on pathognomonic status. As such, determination of its cellular distribution under normal physiological context while utilizing varied techniques is key to illuminating prospective validation of its distinct functions in different tissues. Differential distribution insights at normal physiology would also portent better basis for further clarification of its interactions and proteolytic modifications under pathological conditions like numerous cancer, ischemic stroke and immunomodulation. We therefore raised an antibody against the C terminal of PTMA to use in tandem with available antibody against the N terminal in a murine model to explicate the differences in its distribution in brain cell types and major peripheral organs through western blotting and immunohistochemical approaches.
  
  
  RESULTS
  The newly generated antibody was applied against the N-terminal antibody to distinguish truncated versions of PTMA or deduce possible masking of the protein by other interacting molecules. Western blot analysis indicated presence of a truncated form of the protein only in the thymus, while immunohistochemical analysis showed that in brain hippocampus the full-length PTMA was stained prominently in the nucleus whereas in the stomach full-length PTMA staining was not observed in the nucleus but in the cytoplasm.
  
  
  CONCLUSIONS
  Truncated PTMA could not be detected by western blotting when both antibodies were applied in all tissues examined except the thymus. However, immunohistochemistry revealed differential staining by these antibodies suggesting possible masking of epitopes by interacting molecules. The differential localization patterns observed in the context of nucleic versus cytoplasmic presence as well as punctate versus diffuse pattern in tissues and cell types, warrant further investigations as to the forms of PTMA interacting partners.
authors.affiliation: Department of Molecular Microbiology and Immunology, Division of Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan. kijogi@live.com.|Institute of Tropical Medicine and Infectious Diseases-KEMRI (ITROMID-KEMRI), Nairobi, Kenya. kijogi@live.com.
authors.forename: Caroline Mwendwa
authors.initials: CM
authors.lastname: Kijogi
authors.affiliation: African Population and Health Research Center (APHRC), P. O. Box 10787-00100, Nairobi, Kenya. khayekachris@yahoo.com.|Institute of Tropical Medicine and Infectious Diseases-KEMRI (ITROMID-KEMRI), Nairobi, Kenya. khayekachris@yahoo.com.
authors.forename: Christopher
authors.initials: C
authors.lastname: Khayeka-Wandabwa
authors.affiliation: Department of Pharmacology and Therapeutic Innovation, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan. ksasaki@nagasaki-u.ac.jp.
authors.forename: Keita
authors.initials: K
authors.lastname: Sasaki
authors.affiliation: Department of Pharmacology and Therapeutic Innovation, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan. ystanaka@nagasaki-u.ac.jp.
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Tanaka
authors.affiliation: Department of Pharmacology and Therapeutic Innovation, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan. hkurosu@nagasaki-u.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kurosu
authors.affiliation: Department of Pharmacology and Therapeutic Innovation, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan. hayatom@nagasaki-u.ac.jp.
authors.forename: Hayato
authors.initials: H
authors.lastname: Matsunaga
authors.affiliation: Department of Pharmacology and Therapeutic Innovation, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan. ueda@nagasaki-u.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ueda
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D015153:Blotting, Western / Q000379:methods; D002467:Cell Nucleus / Q000378:metabolism; D005260:Female; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D051379:Mice; D011498:Protein Precursors / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D013947:Thymosin / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism
chemical_list: D000906:Antibodies; D011498:Protein Precursors; C040782:prothymosin alpha; D013947:Thymosin