PMID- 10751034
OWN - NLM
STAT- MEDLINE
DCOM- 20000605
LR  - 20190818
IS  - 0724-8741 (Print)
IS  - 0724-8741 (Linking)
VI  - 17
IP  - 2
DP  - 2000 Feb
TI  - Metal-catalyzed oxidation of brain-derived neurotrophic factor (BDNF): analytical 
      challenges for the identification of modified sites.
PG  - 190-6
AB  - PURPOSE: We examined the metal-catalyzed oxidation of brain-derived neurotrophic 
      factor (BDNF) using the Cu(II)/ascorbate/O2 model oxidative system. METHODS: 
      Electrospray ionization mass spectrometry, peptide mapping and amino acid 
      analysis were utilized to determine the nature of the covalent modification 
      induced by the metal-catalyzed oxidative system. Additionally, analytical 
      ultracentrifugation, the Bradford assay, circular dichroism and ANSA dye-binding 
      were used to determine the nature of any conformational changes induced by the 
      oxidation. RESULTS: Exposure of BDNF to the Cu(II)/ascorbate/O2 system led to the 
      modification of ca. 35% of Met92 to its sulfoxide, and to subsequent 
      conformational changes. The proteolytic digestion procedure was sensitive to this 
      conformational change, and was unable to detect the modification. Chemical 
      digestion with CNBr, however, was not sensitive to this change, and allowed for 
      the identification of the site of modification. CONCLUSIONS: The modification of 
      Met92 to its sulfoxide rendered the oxidized BDNF inaccessible to proteolytic 
      digestion, due to conformational changes associated with the oxidation.
FAU - Jensen, J L
AU  - Jensen JL
AD  - Department of Pharmaceutical Chemistry, University of Kansas, Lawrence 66047, 
      USA.
FAU - Kolvenbach, C
AU  - Kolvenbach C
FAU - Roy, S
AU  - Roy S
FAU - Schoneich, C
AU  - Schoneich C
LA  - eng
GR  - P01AG12993-02/AG/NIA NIH HHS/United States
GR  - T32 GM08359-11/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Pharm Res
JT  - Pharmaceutical research
JID - 8406521
RN  - 0 (Anilino Naphthalenesulfonates)
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Fluorescent Dyes)
RN  - 789U1901C5 (Copper)
RN  - 83662-03-7 (6-anilino-1-naphthalenesulfonic acid)
RN  - AE28F7PNPL (Methionine)
RN  - EC 3.4.- (Endopeptidases)
RN  - OS382OHJ8P (Cyanogen Bromide)
RN  - PQ6CK8PD0R (Ascorbic Acid)
RN  - S88TT14065 (Oxygen)
RN  - XN1XVI4B2C (methionine sulfoxide)
SB  - IM
MH  - Amino Acid Sequence
MH  - Anilino Naphthalenesulfonates/analysis/chemistry/pharmacology
MH  - Ascorbic Acid/metabolism
MH  - Brain-Derived Neurotrophic Factor/analysis/chemistry/*metabolism
MH  - Catalytic Domain
MH  - Chromatography, High Pressure Liquid
MH  - Copper/chemistry/*metabolism
MH  - Cyanogen Bromide/chemistry
MH  - Endopeptidases/pharmacology
MH  - Fluorescent Dyes/analysis/chemistry/pharmacology
MH  - Methionine/analogs & derivatives/chemistry
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - Oxidation-Reduction
MH  - Oxygen/metabolism
MH  - Protein Conformation
MH  - Protein Denaturation
MH  - Sequence Analysis, Protein
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
MH  - Ultracentrifugation
EDAT- 2000/04/06 09:00
MHDA- 2000/06/10 09:00
CRDT- 2000/04/06 09:00
PHST- 2000/04/06 09:00 [pubmed]
PHST- 2000/06/10 09:00 [medline]
PHST- 2000/04/06 09:00 [entrez]
AID - 10.1023/a:1007569431038 [doi]
PST - ppublish
SO  - Pharm Res. 2000 Feb;17(2):190-6. doi: 10.1023/a:1007569431038.