Olink

Olink®
Part of Thermo Fisher Scientific

CARDIOMETABOLIC

Gene
PAM

Uniprot
P19021

Protein
Peptidyl-glycine alpha-amidating monooxygenase

See alternative names Peptidylamidoglycolate lyase

Uniprot Function Description

Bifunctional enzyme that catalyzes the post-translational modification of inactive peptidylglycine precursors to the corresponding bioactive alpha-amidated peptides, a terminal modification in biosynthesis of many neural and endocrine peptides (PubMed:12699694). Alpha-amidation involves two sequential reactions, both of which are catalyzed by separate catalytic domains of the enzyme. The first step, catalyzed by peptidyl alpha-hydroxylating monoxygenase (PHM) domain, is the copper-, ascorbate-, and O2- dependent stereospecific hydroxylation (with S stereochemistry) at the alpha-carbon (C-alpha) of the C-terminal glycine of the peptidylglycine substrate (PubMed:12699694). The second step, catalyzed by the peptidylglycine amidoglycolate lyase (PAL) domain, is the zinc-dependent cleavage of the N-C-alpha bond, producing the alpha-amidated peptide and glyoxylate (PubMed:12699694). Similarly, catalyzes the two-step conversion of an N-fatty acylglycine to a primary fatty acid amide and glyoxylate (By similarity).

Sample type

Human EDTA plasma and serum are the recommended sample types. Human citrate plasma and heparin plasma are also accepted. For other samples types e.g cerebrospinal fluid, (CSF), tissue or cell lysates please we recommend Olink Target 96 panels. Please note that protein expression levels are expected to vary in different sample types and certain assays may be affected by interfering substances such as hemolysate.

Precision

Precision (repeatability) is calculated from linearized NPX values over LOD.

Within run precision Coefficient of Variation (CV)
4%
Between run precision Coefficient of Variation (CV)
6%

Analytical measuring range

The technical data reported below refers to the measured value in the in vitro validation assays run using known concentrations of recombinant antigen. Please note that these figures are for reference only and CANNOT be used to convert NPX values to absolute concentrations for proteins measured in plasma or serum samples.

LOD (pg/mL)
97.7
LLOQ (pg/mL)
195.3
ULOQ (pg/mL)
200000
Hook (pg/mL)
800000
Range (logs)
3

Dilution factor

For optimal assay readout, Olink Explore is run using different dilutions of the original samples (undiluted, 1:10, 1:100 or 1:1000). The dilution factor for this assay is noted below and should be taken into account when estimating the appropriate addressable biological concentration of the protein based on the in vitro validation data.

Dilution factor
1:100

Sensitivity plot

The calibrator curve shown below visualizes the analytical measuring range data based on in vitro measurement of recombinant antigen. Please note that this is shown for reference only and CANNOT be used to convert NPX values to absolute concentrations for proteins measured in plasma or serum samples. The vertical dotted lines represent LLOQ and ULOQ respectively, and the horizontal line indicates the LOD.

10⁻⁶10⁻⁴10⁻²10⁰10²10⁴10⁶10⁸121086420−2−4−6−8pg/mLNPXReplicate4PLLODLLOQULOQ

Sample distribution plot

The plot below shows the levels of protein measured in a number of commercial plasma samples. Healthy subjects are shown in blue and samples obtained from patients with a range of diseases are shown in red. The latter include inflammatory, cardiovascular, autoimmune & neurological diseases, as well as cancer. The data is shown to give a general idea of the sort of data range to expect, but cannot cover all potential levels that may be seen in clinical samples.

121086420−2−4−6−8NPXControl SamplesDisease Samples