SUMMARY
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.
KEY WORDS
[G14]-humanin; humanin derivatives; MALDI-TOF MS; PSD; peptide oxidation
REFERENCES
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, www.siumed.edu/neuro/cadrd.html; 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.
|
CITED
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.
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