dba

 

Lysyl Endopeptidase®

Lysyl endopeptidase is a serine protease that cleaves peptide bonds at the carboxy-terminal side of lysine residues with a high degree of specificity. It is inhibited by TLCK, PMSF, and DFP but not by TACK. Taking advantage of its excellent specificity, lysyl endopeptidase is used for peptide fragmentation and peptide mapping for analysis of the primary structure of proteins.

It is originally isolated from a soil bacterium by Masaki et al., cleaves specifically the peptides on the carboxyterminal side of Lysine residues. This enzyme is very useful both in protein sequence analysis and in enzymatic synthesis of Lys-X compounds.

  • Molecular weight: 27,000(Gel filtration), 30,000(SDS-PAGE)
  • Optimal pH: 9.0~9.5(Amidase activity)
  • Isoelectric point: 6.9~7.0
  • Substrate specificity
           -Hydrolysable substrates???Tos-Lys-OMe, Bz-Lys-NH2, Bz-Lys-pNA, Lys-pNA
           -Unhydrolysable substrates???Bz-Arg-NH2, Bz-Arg-pNA, Arg-pNA
  • Inhibitors: DFP, PMSF, TLCK

    Available as Research (Biochemistry) or Mass Spectrometry Grade

     

    Product Number

    Product name

    Package Size

    129-02541

    Lysyl EndopeptidaseR (Lys-C)

    for Biochemistry

     

    1vial(10AU)

    125-02543

    Lysyl EndopeptidaseR (Lys-C)

    for Biochemistry

     

    1vial(2AU)

    125-05061

    Lysyl Endopeptidase, Mass Spectrometry Grade (Lys-C)

    20ug x5

    Comparison of In-gel Digestion using Trypsin (Tp), Lysl Endopeptidase (Lep) and Lep Combined with Tp (Lep + Tp)

    BSA band (100 ng) resolved by SDS-PAGE was in-gel digested with Tp, Lep and Lep + Tp and analyzed by MALDI-TOFMAS. The figure shows the individual mass spectra. The evaluation of these peptidases is summarized in the table.

    Mass spectrum using Tp and Tp + Lep

    00062325_img01.png

    When Tp is used concurrently with Lep, additional peaks were found around m/z 2000 compared to when only Tp is used. It shows that the recovery rate of peptide increases.

    Table. Comparison ofTp, Lep, and Tp + Lep

    Tp Lep Tp + Lep
    Cleavage site C-terminal of Arg and Lys C-terminal of Lys C-terminal of Arg and Lys
    Missed Cleavage(Rates of missed cleavage)? Many
    (8 %)    		 Very few
    (0 %)    		 Few
    (3 %)
    Number of identified peptides 17 19 22

    ?The coverage (percentage of available peptides in the entire sequence) obtained by database search using a parameter setting of Missed Cleavage = 0 was subtracted from the coverage obtained by using a parameter setting of Missed Cleavage = 1.The value resulted from subtracting the coverage obtained when database searches were performed with Missed cleavage 0 from that obtained when performed with Missed cleavage 1. “Coverage” is the percentage of peptides obtained after in-gel digestion in the whole sequence.

    These results indicate there are very few missed cleavages obtained by Lep digestion. When Tp is used concurrently with Lep, missed cleavages decrease and the number of identified peptides increase compared to when only Tp is used.


    •  

    Richiedi un'offerta

    DBA-ITALIA Srl
    Via Umbria 10
    20090 Segrate (Milano)

    Tel. +39 02-26922300
    Fax +39 02-26926058
    Fax +39 02-26923535




    If you no longer wish to receive these announcements click here