title: Identification and characterization of neutral endopeptidase (EC 3. 4. 24. 11) from human prostasomes--localization in prostatic tissue and cell lines.
abstract: BACKGROUND
  An antibody directed against a 100 kDa protein was immunoselected from a polyvalent antiserum against human prostasomes. The antibody as well as biochemical characteristics of the respective antigen were used to study the structural relationship of the latter with prostate membrane specific antigen (PMSA), another 100 kDa membrane protein of the prostate.
  
  
  METHODS
  The isolated purified 100 kDa protein was characterized by tryptic degradation, aminoacid-sequencing and mass spectroscopy peptide-fingerprinting as well as mono-saccharide analysis and lectin binding and identified as a prostasomal neutral endopeptidase (NEP, EC 3.4.24.11). Immunohistochemistry, immunoelectron microscopy, in situ hybridization, and RT-PCR were performed to analyze the expression and distribution of the protein in normal and malignant human prostatic tissues and cell lines.
  
  
  RESULTS
  Prostatic NEP, which has no relationship with PMSA, is a glycosylated, integral membrane protein type II. The prevalent glycosyl residues are NeuNAc, GlcNAc, GalNAc, Gal, Man, Fuc. NEP-mRNA is expressed in human prostatic epithelial and some stromal cells. NEP-immunoreactivity is strong in normal prostatic epithelium and confined to the apical plasma membrane. During apocrine secretion, the enzyme is released from the secretory cells, contributing to the formation of prostasomes. In prostate cancer specimens, immunoreactivity of apical plasma membranes is lost, while generalized cytoplasmic immunoreactivity develops.
  
  
  CONCLUSIONS
  Prostatic secretory cells contain a membrane-bound, highly glycosylated neutral endopeptidase which is restricted to the apical plasma membrane. The enzyme is released from the cells in an apocrine fashion and contributes to the formation of prostasomes. In prostate cancer cells a preferential cytoplasmic localization is observed, pointing to alterations in intracellular targeting.
authors.affiliation: Department of Urology, Municipal Hospital, Offenbach, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Renneberg
authors.forename: M
authors.initials: M
authors.lastname: Albrecht
authors.forename: R
authors.initials: R
authors.lastname: Kurek
authors.forename: E
authors.initials: E
authors.lastname: Krause
authors.forename: F
authors.initials: F
authors.lastname: Lottspeich
authors.forename: G
authors.initials: G
authors.lastname: Aumüller
authors.forename: B
authors.initials: B
authors.lastname: Wilhelm
mesh_terms: D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D002460:Cell Line; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D015260:Neprilysin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011467:Prostate / Q000201:enzymology* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D022142:Secretory Vesicles / Q000201:enzymology*; D012661:Semen / Q000201:enzymology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: D012333:RNA, Messenger; D015260:Neprilysin