Sequence proteins and protein segments inaccessible with trypsin
See what you are missing. Additional sequence coverage and ID's.
Difficult Proteins? Maybe you need a better Protease.
HTA-Proteases provide additional sequence coverage and protein identifications with datasets orthogonal to trypsin and other available enzymes. Within one hour you can be running your HTA-samples on your LC-MS/MS.
Rapid and simple Histone sequencing
Obtain histone coverage from whole-cell extracts with NO chemical treatments or manipulation. Less than one hour from raw sample to histone sequence data. The cleavage preference of HTA-Proteases (E, L, F) obviates chemical blocking of lysine that is required for tryptic approaches to sequence histones and other proteins with abundant lysine and arginine residues.
HTA-Proteases give additional sequence for >50% of tryptic identifications with over 100 additional non-tryptic identifications
Commercial K562 extract was digested in parallel with trypsin or Krakatoa or Vesuvius. 53% of the identified proteins showed improved coverage with inclusion of HTA-Protease data. 285 proteins showed sequence coverage increases by 20% or more relative to trypsin alone with inclusion of the HTA-Protease data. Strikingly, 52% of the proteins with >20% additional coverage were nucleic acid-binding proteins. Of the same 285 proteins, 14% were identified as mitochondrial proteins, most of which are membrane proteins, with an additional 13% of these proteins annotated as either chaperones or nuclear proteins. Taken together, 79% of the proteins with 20% or more increased coverage classified into these four categories.