ISSN 1214-0287 (on-line)
ISSN 1214-021X (printed)

Volume 2 (2004), No 2, p 113-117

MALDI-TOF mass spectrometric properties of humanin-like peptides

Klara Novotna, Ondrej Sedo, Josef Havel

Address: Josef Havel, Department of Analytical Chemistry, Faculty of Science, Masaryk University, Kotlarska 2, 611 37 Brno, Czech Republic

Received 21st January 2004.
Revised 19th February 2004.
Published online 16th April 2004.

Full text article (pdf)

The possibilities of MALDI-TOF mass spectrometric analysis of new neuro-protective peptide [G14]-humanin (HNG) and similar compounds are studied with the aim of finding optimal conditions for the determination of these peptides. Acidification and washing of HNG samples using 5% (v/v) formic acid is necessary to reach a detection limit similar to other peptides. The sensitivity of HNG determination is decreased in the oxidative environment as the peptide yields oxidation of methionine and cysteine forming several species, including a disulfide dimer. During Post-source Decay (PSD) it was found that intense cleavage between Asp and Leu in HNG reduces the possibility of detecting other fragments. Better sequence coverage is gained from shorter humaninlike peptides.

[G14]-humanin; humanin derivatives; MALDI-TOF MS; PSD; peptide oxidation

Chaurand P, Luetzenkirchen F, Spengler BJ: Peptide and protein identification by matrixassisted laser desorption ionization (MALDI) and MALDI-post-source decay time-of-flight mass spectrometry. Am Mass Spectrom 10:91-103, 1999.

Godovac-Zimmermann J, Brown LR: Perspectives for mass spectrometry and functional proteomics. Mass Spectrom Rev 20:1-57, 2001.

Guo B, Zhai D, Cabezas E, Welsh K, Nouraini S, Satterthwait AC, Reed JC: Humanin peptide suppresses apoptosis by interfering with Bax activation. Nature 423:456-461, 2003.

Hashimoto Y, Ito Y, Niikura T, Shao Z, Hata M, Oyama F, Nishimoto I: Mechanisms of neuro-protection by novel rescue factor humanin from Swedish mutant amyloid precursor protein. Biochem Biophys Res Commun 283:460-468, 2001.

Ikonen M, Liu B, Hashimoto Y, Ma L, Lee KW, Niikura T, Nishimoto I, Cohen P: Interaction between the Alzheimer's survival peptide humanin and insulin-like growth factor-binding protein 3 regulates cell survival and apoptosis. Proc Natl Acad Sci USA 100:13042-13047, 2003.

Jung SS, Van Nostrand WEJ: Humanin rescues human cerebrovascular smooth muscle cells from Abeta-induced toxicity. Neurochem 84:266-272, 2003.

Kaufmann R, Kirsch D, Spengler B: Sequencing of peptides in a time-of-flight mass spectrometer: evaluation of post-source decay following matrix-assisted laser desorption ionization (MALDI). Int J Mass Spectrom Ion Processes 131:355-385, 1994.

Reiber D, Grover TA, Brown RS: Identifying protein using matrix-assisted laser desorption/ionization in-source fragmentation data combined with database searching. Anal Chem 70:673-683, 1998.

Sedo O, Novotna K, Havel J: Rapid Commun Mass Spectrom, in press.

Terashita K, Hashimoto Y, Niikira T, Tajima H, Yamagishi Y, Ishizaka M, Kawasumi M, Chiba T, Kanekura K, Yamada M, Nawa M, Kita Y, Aiso S, Nishimoto I: Two serine residues distinctly regulate the rescue function of Humanin, an inhibiting factor of Alzheimer's disease-related neurotoxicity: functional potentiation by isomerization and dimerization. J Neurochem 85, 1521-1538,; 2003.

Yamagishi Y, Hashimoto Y, Niikura T, Nishimoto I: Identification of essential amino acids in Humanin, a neuroprotective factor against Alzheimer's disease-relevant insults. Peptides 24:585-595, 2003.

Zou P, Ding Y, Sha Y, Hu B, Nie S: Humanin peptides block calcium influx of rat hippocampal neurons by altering fibrogenesis of Abeta (1-40). Peptides 24:679-685, 2003.

Novotna K, Havlis J, Havel J: HPLC separation-of neuroprotective peptides - General approach, towards optimization: experimental sesign combined with artificial neural networks. Pept 2004, Proceedings - Bridges Between Disciplines 469-470, 2005.

Novotna K, Havlis J, Havel J: Optimisation of high performance liquid chromatography separation of neuroprotective peptides - Fractional experimental designs combined with artificial neural networks. J Chromatogr A 1096:50-57, 2005.