FEBS Letters | |
The active site region of the vitamin K‐dependent carboxylase includes both the amino‐terminal hydrophobic and carboxy‐terminal hydrophilic domains of the protein | |
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
【 预 览 】
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