Direct analysis of α- and β-chains of hemoglobins from mammalian blood samples by nanoESI mass spectrometry during in-capillary proteolytic digestion

Henning, Stefanie; Mormann, Michael; Peter-Katalinić, Jasna; Pohlentz, Gottfried
July 2011
Amino Acids;Jul2011, Vol. 41 Issue 2, p343
Academic Journal
α- and β-chains of hemoglobins derived from several species were analyzed directly from diluted blood samples by simultaneous in-capillary proteolytic digestion and nanoESI MS and MS/MS analysis. Starting from fresh or frozen and thawed blood samples, sequence coverages of >80% were usually obtained. Only 2 h after resuspension of a dried blood spot, human origin could be demonstrated from data obtained by in-capillary tryptic digestion, nanoESI mass spectrometric analysis, and data base search. A fast and facile differentiation of closely related species by hemoglobin-derived proteolytic 'marker peptides' was demonstrated for Asian ( Elephas maximus) and African elephants ( Loxodonta africana). Finally, amino acid sequences deduced from collision-induced dissociation experiments during in-capillary proteolytic digestion of the corresponding blood samples allowed de novo sequencing of previously unknown sequences of hemoglobin chains of the Patagonian cavy ( Dolichotum patagona) and the Persian gazelle ( Gazella subgutturosa subgutturosa). 100% of the α-chain sequences and more than 85% of the β-chain sequences were covered for both the species. Additionally, sequence data derived from tandem MS experiments obtained with the Q-Tof analyzer were confirmed by high resolution Fourier-transform ion cyclotron resonance mass spectrometric experiments. Accurate protein mass determination of the intact hemoglobin chains directly from the corresponding blood samples by use of a Fourier-transform ion cyclotron resonance mass spectrometer corroborated the deduced sequences of the respective α-chains. The present study demonstrates that in-capillary digestion allows fast characterization and/or sequencing of hemoglobin chains directly from blood samples.


Related Articles

  • CSTX-9, a toxic peptide from the spider Cupiennius salei: amino acid sequence, disulphide bridge pattern and comparison with other spider toxins containing the cystine knot structure. Schaller, J.; Kämpfer, U.; Schürch, S.; Kuhn-Nentwig, L.; Haeberli, S.; Nentwig, W. // Cellular & Molecular Life Sciences;Sep2001, Vol. 58 Issue 10, p1538 

    CSTX-9 (68 residues, 7530.9 Da) is one of the most abundant toxic polypeptides in the venom of the wandering spider Cupiennius salei. The amino acid sequence was determined by Edman degradation using reduced and alkylated CSTX-9 and peptides generated by cleavages with endoproteinase Asp-N and...

  • Interrogating the Venom of the Viperid Snake Sistrurus catenatus edwardsii by a Combined Approach of Electrospray and MALDI Mass Spectrometry. Chapeaurouge, Alex; Reza, Md Abu; Mackessy, Stephen P.; Carvalho, Paulo C.; Valente, Richard H.; Teixeira-Ferreira, André; Perales, Jonas; Lin, Qingsong; Kini, R. Manjunatha // PLoS ONE;May2015, Vol. 10 Issue 5, p1 

    The complete sequence characterization of snake venom proteins by mass spectrometry is rather challenging due to the presence of multiple isoforms from different protein families. In the present study, we investigated the tryptic digest of the venom of the viperid snake Sistrurus catenatus...

  • A Trunk Full of Surprises. Searl, Duncan // Elephants (1-59716-162-4);2006, p10 

    Information on elephant trunk from a chapter of the book "Elephants" is presented. The nostrils of elephants are located at the tip of their trunks. Elephants use their trunks mainly for eating and drinking. The two kinds of elephants, Asian and African, are distinguished by the tip of their trunks.

  • Chemical Signals of Elephant Musth: Temporal Aspects of Microbially-Mediated Modifications. Goodwin, Thomas; Broederdorf, Laura; Burkert, Blake; Hirwa, Innocent; Mark, Daniel; Waldrip, Zach; Kopper, Randall; Sutherland, Mark; Freeman, Elizabeth; Hollister-Smith, Julie; Schulte, Bruce // Journal of Chemical Ecology;Jan2012, Vol. 38 Issue 1, p81 

    Mature male African ( Loxodonta africana) and Asian ( Elephas maximus) elephants exhibit periodic episodes of musth, a state in which serum androgens are elevated, food intake typically decreases, aggressiveness often increases, and breeding success is enhanced. Urine is a common source of...

  • Name That Elephant!  // Weekly Reader News - Edition 3;9/9/2005, Vol. 75 Issue 1, p4 

    Presents a quiz about African elephants and Asian elephants.

  • World of Wonder ELEPHANTS.  // Adirondack Daily Enterprise;7/17/2010, Vol. 117 Issue 168, pA6 

    The article offers information the difference between the African elephant and an Asian elephant.

  • “Pseudoalteromonas januaria” SUT 11 as the Source of Rare Lipodepsipeptides. Kalinovskaya, Nataliya I.; Dmitrenok, Andrey S.; Kuznetsova, Tatyana A.; Frolova, Galina M.; Christen, Richard; Laatsch, Hartmut; Alexeeva, Yulia V.; Ivanova, Elena P. // Current Microbiology;Mar2008, Vol. 56 Issue 3, p199 

    The marine bacterium “Pseudoalteromonas januaria” SUT 11 isolated from a seawater sample produced the rare cell-bound cyclic lipodepsipeptides A/A′, B/B′, and C/C′. The matrix-assisted laser desorption/ionization mass spectra indicated that one bromine atom...

  • Synthesis of phytochelatins in vetiver grass upon lead exposure in the presence of phosphorus. Andra, Syam S.; Datta, Rupali; Sarkar, Dibyendu; Makris, Konstantinos C.; Mullens, Conor P.; Sahi, Shivendra V.; Bach, Stephan B. H. // Plant & Soil;Jan2010, Vol. 326 Issue 1/2, p171 

    In a hydroponic setting, we investigated the possible role of phytochelatins (metal-binding peptides) in the lead (Pb) tolerance of vetiver grass ( Vetiveria zizanioides L.). Pb was added to the nutrient medium at concentrations ranging from 0 to 1,200 mg L−1. Furthermore, we simulated the...

  • The Quantitative Analysis of Paraquat in Biological Samples by Liquid Chromatography- Electrospray Ionization-Mass Spectrometry. Zhaohong Wang; Zhiping Wang; Junbo Xing // Journal of Analytical Toxicology;Jan/Feb2011, Vol. 35 Issue 1, p23 

    The article presents a study which examines the use of liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) to detect paraquat distribution in blood and tissue samples. It evaluates five cases of suspected paraquat intoxication, which blood and urine samples were analyzed...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics