PMID- 23303182
OWN - NLM
STAT- MEDLINE
DCOM- 20130509
LR  - 20211021
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 288
IP  - 9
DP  - 2013 Mar 1
TI  - A novel transition-state analogue for lysozyme, 4-O-beta-tri-N-acetylchitotriosyl 
      moranoline, provided evidence supporting the covalent glycosyl-enzyme 
      intermediate.
PG  - 6072-82
LID - 10.1074/jbc.M112.439281 [doi]
AB  - 4-O-beta-Di-N-acetylchitobiosyl moranoline (2) and 4-O-beta-tri-N-acetylchitotriosyl 
      moranoline (3) were produced by lysozyme-mediated transglycosylation from the 
      substrates tetra-N-acetylchitotetraose, (GlcNAc)4, and moranoline, and the 
      binding modes of 2 and 3 to hen egg white lysozyme (HEWL) was examined by 
      inhibition kinetics, isothermal titration calorimetry (ITC), and x-ray 
      crystallography. Compounds 2 and 3 specifically bound to HEWL, acting as 
      competitive inhibitors with Ki values of 2.01 x 10(-5) and 1.84 x 10(-6) m, 
      respectively. From ITC analysis, the binding of 3 was found to be driven by 
      favorable enthalpy change (DeltaHr degrees ), which is similar to those obtained for 2 and 
      (GlcNAc)4. However, the entropy loss (-TDeltaSr degrees ) for the binding of 3 was smaller 
      than those of 2 and (GlcNAc)4. Thus the binding of 3 was found to be more 
      favorable than those of the others. Judging from the Kd value of 3 (760 nm), the 
      compound appears to have the highest affinity among the lysozyme inhibitors 
      identified to date. X-ray crystal structure of HEWL in a complex with 3 showed 
      that compound 3 binds to subsites -4 to -1 and the moranoline moiety adopts an 
      undistorted (4)C1 chair conformation almost overlapping with the -1 sugar 
      covalently bound to Asp-52 of HEWL (Vocadlo, Davies, G. J., Laine, R., and 
      Withers, S. G. (2001) Nature 412, 835-838). From these results, we concluded that 
      compound 3 serves as a transition-state analogue for lysozyme providing 
      additional evidence supporting the covalent glycosyl-enzyme intermediate in the 
      catalytic reaction.
FAU - Ogata, Makoto
AU  - Ogata M
AD  - Department of Bioscience, Graduate School of Science and Technology, Shizuoka 
      University, Shizuoka 422-8529, Japan.
FAU - Umemoto, Naoyuki
AU  - Umemoto N
FAU - Ohnuma, Takayuki
AU  - Ohnuma T
FAU - Numata, Tomoyuki
AU  - Numata T
FAU - Suzuki, Akari
AU  - Suzuki A
FAU - Usui, Taichi
AU  - Usui T
FAU - Fukamizo, Tamo
AU  - Fukamizo T
LA  - eng
SI  - PDB/4HP0
SI  - PDB/4HPI
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20130109
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Enzyme Inhibitors)
RN  - 19130-96-2 (1-Deoxynojirimycin)
RN  - EC 3.2.1.17 (Muramidase)
SB  - IM
MH  - 1-Deoxynojirimycin/*analogs & derivatives/*chemistry
MH  - Animals
MH  - Catalysis
MH  - Chickens
MH  - Crystallography, X-Ray
MH  - Enzyme Inhibitors/*chemistry
MH  - Muramidase/antagonists & inhibitors/*chemistry/metabolism
MH  - Protein Binding
MH  - Protein Structure, Tertiary
PMC - PMC3585046
EDAT- 2013/01/11 06:00
MHDA- 2013/05/10 06:00
PMCR- 2014/03/01
CRDT- 2013/01/11 06:00
PHST- 2013/01/11 06:00 [entrez]
PHST- 2013/01/11 06:00 [pubmed]
PHST- 2013/05/10 06:00 [medline]
PHST- 2014/03/01 00:00 [pmc-release]
AID - S0021-9258(20)43502-1 [pii]
AID - M112.439281 [pii]
AID - 10.1074/jbc.M112.439281 [doi]
PST - ppublish
SO  - J Biol Chem. 2013 Mar 1;288(9):6072-82. doi: 10.1074/jbc.M112.439281. Epub 2013 
      Jan 9.