Direct analysis of the products of sequential cleavages of peptides and proteins affinity-bound to immobilized metal ion beads by matrix-assisted laser desorption/ionization mass spectrometry.
Consecutive enzymatic reactions on analytes affinity-bound to immobilized metal ion beads with subsequent direct analysis of the products by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry have been used for detecting protein synthesis errors occuring at the N-terminus. The usefulness of this method was demonstrated by analyzing two commercially available recombinant HIV proteins with affinity tags at the N-terminus, and histatin-5, a peptide with multiple histidine residues. The high specificity, sensitivity, and speed of analysis make this method especially useful in obtaining N-terminal sequencing information of histidine-tagged recombinant proteins.
Qian
X
X
Laboratory of Structure Biology, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, 27709, USA.
Zhou
W
W
Khaledi
M G
MG
Tomer
K B
KB
D000595:Amino Acid Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D016346:Gene Products, vif / Q000032:analysis; D016655:HIV Core Protein p24 / Q000032:analysis; D015497:HIV-1; D008670:Metals; D008969:Molecular Sequence Data; D011506:Proteins / Q000032:analysis; D011994:Recombinant Proteins / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014764:Viral Proteins / Q000032:analysis*; D054320:vif Gene Products, Human Immunodeficiency Virus; D000099053:Histidine-Rich Glycoprotein
D016346:Gene Products, vif; D016655:HIV Core Protein p24; D008670:Metals; D011506:Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D000099053:Histidine-Rich Glycoprotein; D054320:vif Gene Products, Human Immunodeficiency Virus