title: Genetic determinants of amidating enzyme activity and its relationship with metal cofactors in human serum.
abstract: BACKGROUND
  α-amidation is a final, essential step in the biosynthesis of about half of all peptide hormones and neurotransmitters. Peptidylglycine α-amidating monooxygenase (PAM), with enzymatic domains that utilize Cu and Zn, is the only enzyme that catalyzes this reaction. PAM activity is detected in serum, but its significance and utility as a clinical biomarker remain unexplored.
  
  
  METHODS
  We used well-established enzymatic assays specific for the peptidylglycine-α -hydroxylating monooxygenase (PHM) and peptidyl-α-hydroxyglycine α-amidating lyase (PAL) domains of PAM to quantify amidating activity in the sera of 144 elderly men. Relationships between PHM and PAL activity and serum levels of their respective active-site metals, Cu and Zn, were analyzed. Study participants were also genotyped for eight non-coding single nucleotide polymorphisms (SNPs) in PAM, and relationships between genotype and serum enzyme activity and metal levels were analyzed.
  
  
  RESULTS
  Serum PHM and PAL activities were normally distributed and correlated linearly with each other. Serum PAL activity, but not serum PHM activity, correlated with serum Cu; neither activity correlated with serum Zn. Study subjects possessing the minor alleles for rs32680 had lower PHM and PAL activities, and subjects with minor alleles for rs11952361 and rs10515341 had lower PHM activities.
  
  
  CONCLUSIONS
  Our results characterize large variation in serum amidating activity and provide unique insight into its potential origin and determinants. Common non-coding polymorphisms affect serum amidating activity and Cu levels. Serum amidating activity should be explored as a biomarker for functionality in the elderly and in additional study groups.
authors.forename: Eric D
authors.initials: ED
authors.lastname: Gaier
authors.forename: Alison
authors.initials: A
authors.lastname: Kleppinger
authors.forename: Martina
authors.initials: M
authors.lastname: Ralle
authors.forename: Jonathan
authors.initials: J
authors.lastname: Covault
authors.forename: Richard E
authors.initials: RE
authors.lastname: Mains
authors.forename: Anne M
authors.initials: AM
authors.lastname: Kenny
authors.affiliation: Department of Neuroscience, University of Connecticut Health Center, 06030 Farmington, CT, USA. eipper@uchc.edu.
authors.forename: Betty A
authors.initials: BA
authors.lastname: Eipper
mesh_terms: D000368:Aged; D003300:Copper / Q000097:blood*; D004247:DNA / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D015032:Zinc / Q000097:blood*
chemical_list: D009097:Multienzyme Complexes; D003300:Copper; D004247:DNA; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D015032:Zinc