The active site region of the vitamin K‐dependent carboxylase includes both the amino‐terminal hydrophobic and carboxy‐terminal hydrophilic domains of the protein

FEBS Letters

Maillet, Magali²  Marquet, André²  Morris, Daniel¹  Gaudry, Michel² 

[1] Department of Biology, CB# 3280, Coker Hall, University of North Carolina, Chapel Hill, NC 27599, USA;Laboratoire de Chimie Organique Biologique, Université Pierre et Marie Curie, Tour 44–45, 4 place Jussieu, 75252 Paris Cedex 05, France

关键词: Vitamin K-dependent carboxylase;    Photolabeling;    p-Benzoylphenylalanine;    Enzyme inactivation;    Catalytic site;    Propeptide binding site;    Bpa;    p-benzoylphenylalanine;    CHAPS;    3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate;    CP1 or 2;    carboxylase probe 1 or 2 (see Table 1);    kDaapp;    apparent mass in kDa on SDS-PAGE;    MOPS;    4-morpholinopropanesulfonic acid;    NαBoc;    αNH2 protected by terbutoxycarbonyl group;    PVDF;    polyvinylidene fluorure;    TCA;    trichloroacetic acid;   

DOI  :  10.1016/S0014-5793(97)00831-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In order to localize the active site of the vitamin K-dependent carboxylase, we developed an affinity probe containing the propeptide and the first two carboxylatable glutamate residues conserved in many native substrates. This probe crosslinked to both the hydrophobic amino-terminal and hydrophilic carboxy-terminal domains of the carboxylase, in contrast with previous work which localized both the catalytic and the propeptide binding site within the amino-terminal hydrophobic domain. Amino acid analysis revealed that the mass of an amino-terminal fragment is seriously underestimated by SDS-PAGE. Reanalysis of the published data in light of this information suggests that a portion of the propeptide binding site resides within the carboxy-terminal hydrophilic domain.

【 授权许可】

Unknown