PMID- 10502668
OWN - NLM
STAT- MEDLINE
DCOM- 19991230
LR  - 20190513
IS  - 0021-924X (Print)
IS  - 0021-924X (Linking)
VI  - 126
IP  - 4
DP  - 1999 Oct
TI  - Identification of cysteine-380 as the essential residue for the human 
      N-acetyl-D-glucosamine 2-epimerase (renin binding protein).
PG  - 639-42
AB  - Renin binding protein (RnBP) is a proteinous renin inhibitor firstly isolated 
      from porcine kidney. Recently, the protein was identified as the enzyme, 
      N-acetyl-D-glucosamine (GlcNAc) 2-epimerase. The GlcNAc 2-epimerase activity of 
      recombinant human RnBP was specifically inhibited by SH-reagents such as 
      N-ethylmaleimide, 5, 5'-dithiobis-2-nitrobenzoate, and iodoacetic acid, 
      indicating that the most probable reactive site is a cysteine residue. To 
      identify the active site residue(s), we have constructed ten cysteine residue 
      mutants (C41S, C66S, C104S, C125S, C210S, C239S, C302S, C380S, C386S, and C390S) 
      for human GlcNAc 2-epimerase and expressed them in Escherichia coli cells. The 
      relative specific activities of C41S, C66S, C125S, C210S, C239S, C302S, C386S, 
      and C390S are nearly the same to that of the wild-type enzyme. The specific 
      activity of the C104S mutant is 26% of that of the wild-type enzyme. The 
      expression of the C380S mutant in E. coli cells was detected on Western blotting, 
      whereas GlcNAc 2-epimerase activity was not detected in the extract. These 
      results indicate that Cys380 is essential for the enzymatic activity of human 
      GlcNAc 2-epimerase.
FAU - Takahashi, S
AU  - Takahashi S
AD  - Department of Bioengineering, Akita Research Institute of Food and Brewing 
      (ARIF), Arayamachi, Akita, 010-1623, Japan. saori@arif.pref. akita.jp
FAU - Takahashi, K
AU  - Takahashi K
FAU - Kaneko, T
AU  - Kaneko T
FAU - Ogasawara, H
AU  - Ogasawara H
FAU - Shindo, S
AU  - Shindo S
FAU - Saito, K
AU  - Saito K
FAU - Kobayashi, M
AU  - Kobayashi M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Biochem
JT  - Journal of biochemistry
JID - 0376600
RN  - 0 (Carrier Proteins)
RN  - 0 (DNA Primers)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Sulfhydryl Reagents)
RN  - EC 5.1.3.- (Carbohydrate Epimerases)
RN  - EC 5.1.3.8 (N-acyl-D-glucosamine 2-epimerase)
RN  - EC 5.1.3.8 (RENBP protein, human)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Carbohydrate Epimerases/antagonists & inhibitors/*chemistry/genetics
MH  - Carrier Proteins/antagonists & inhibitors/*chemistry/genetics
MH  - Catalytic Domain/genetics
MH  - Cysteine/chemistry
MH  - DNA Primers/genetics
MH  - Escherichia coli/genetics
MH  - Humans
MH  - Mutagenesis, Site-Directed
MH  - Recombinant Proteins/antagonists & inhibitors/chemistry/genetics
MH  - Sulfhydryl Reagents/pharmacology
EDAT- 1999/09/30 00:00
MHDA- 1999/09/30 00:01
CRDT- 1999/09/30 00:00
PHST- 1999/09/30 00:00 [pubmed]
PHST- 1999/09/30 00:01 [medline]
PHST- 1999/09/30 00:00 [entrez]
AID - 10.1093/oxfordjournals.jbchem.a022496 [doi]
PST - ppublish
SO  - J Biochem. 1999 Oct;126(4):639-42. doi: 10.1093/oxfordjournals.jbchem.a022496.