Isolation and characterization of serum procalcitonin from patients with sepsis.
Procalcitonin (PCT) is one of the precursors in the synthesis of calcitonin in thyroidal C-cells and other neuroendocrine cells. PCT and other calcitonin precursors are elevated in the serum of many conditions leading to systemic inflammatory response syndrome. The measurement of PCT in patients suffering from severe bacterial infections is a useful tool for the diagnosis of sepsis. Furthermore, therapeutic decisions are often based on the increase or decline of serum PCT levels. PCT was reported to have 116 amino acids. The aim of our study was the determination of the primary structure of serum PCT from septic patients. Sera containing high PCT-concentrations (>100 ng/ml) were collected from 22 patients with severe sepsis and were pooled for further purification (12.7 microg total concentration of PCT). Pooled PCT was purified on a CT 21-immunoaffinity column, further purified by reversed phase HPLC, and the resulting pure PCT was digested with endoproteinase Asp-N. N-terminal Edman sequencing showed that the first two amino acids (Ala-Pro) of the proposed pro-peptide were missing. Further analyses by MALDI-TOF mass spectroscopy resulted in a distinct mass signal of 12640 Da +/- 0.1%, which is in concordance with the theoretical molecular weight of the N-terminal truncated form (12628 Da). As opposed to previous suggestions, we could not detect any chemical modifications of PCT. In summary, we could demonstrate that PCT in the serum of septic patients is a peptide of only 114 amino acids, instead of the predicted 116 amino acids, lacking the N-terminal dipeptide Ala-Pro. This information on the primary structure of PCT might help in further studies on the physiological role of PCT during sepsis.
Weglöhner
W
W
InVivo Diagnostica Entwicklungsgesellschaft mbH, Neuendorfstr. 25, D-16761, Hennigsdorf, Germany. j.struck@brahms.de
Struck
J
J
Fischer-Schulz
C
C
Morgenthaler
N G
NG
Otto
A
A
Bohuon
C
C
Bergmann
A
A
D000595:Amino Acid Sequence; D002116:Calcitonin / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D015740:Calcitonin Gene-Related Peptide; D002846:Chromatography, Affinity; D006801:Humans; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011498:Protein Precursors / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D018805:Sepsis / Q000097:blood*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
C000606589:CALCA protein, human; D011498:Protein Precursors; D002116:Calcitonin; D015740:Calcitonin Gene-Related Peptide