The detection of nitrated tyrosine in neuropeptides: a MALDI matrix-dependent response.
Neuropeptides are a diverse class of signaling molecules that typically have one or more posttranslational modifications. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is an effective tool for identification and characterization of neuropeptides from samples as small as individual neurons. However, the detection of one particular posttranslational modification-nitrotyrosine-has been problematic because of the lability of the nitro group of nitrotyrosine under MALDI-MS conditions. The detection of nitrated tyrosine in peptide standards was dependent on the MALDI matrix used for the analysis. Specifically, sinapinic acid was the optimum matrix tested to observe this modification while it was not consistently detected with matrices such as 2,5-dihydroxybenzoic acid. Using the optimized procedures, several identified nitric-oxide-synthase positive neurons from Lymnaea stagnalis were tested to determine if the neuropeptides present were nitrated. In all cases, the nitrated form of the neuropeptide was not observed. The dependence on the sample-preparation procedures of observing this particular chemical modification demonstrates the need for careful selection of sample-preparation methods with MALDI or the use of other ionization methods.
Sheeley
Sarah A
SA
Department of Chemistry and the Beckman Institute, University of Illinois at Urbana-Champaign, 600 S. Mathews Ave 63-5, Urbana, IL 61801, USA.
Rubakhin
Stanislav S
SS
Sweedler
Jonathan V
JV
D000595:Amino Acid Sequence; D000818:Animals; D001048:Aplysia; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000737:chemistry*; D009566:Nitrates / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014443:Tyrosine / Q000032:analysis* / Q000737:chemistry
D009479:Neuropeptides; D009566:Nitrates; D014443:Tyrosine