PMID- 8251941
OWN - NLM
STAT- MEDLINE
DCOM- 19940113
LR  - 20181113
IS  - 0961-8368 (Print)
IS  - 1469-896X (Electronic)
IS  - 0961-8368 (Linking)
VI  - 2
IP  - 10
DP  - 1993 Oct
TI  - Sites of iodination in recombinant human brain-derived neurotrophic factor and 
      its effect on neurotrophic activity.
PG  - 1664-74
AB  - Recombinant human brain-derived neurotrophic factor (BDNF) is now under extensive 
      investigation because of its potential clinical applications. Radioactively 
      labeled proteins are usually required to study receptor binding and 
      pharmacokinetic properties of proteins. This study was undertaken to see if 
      iodination affects the biological and conformational properties of a recombinant 
      BDNF. BDNF was iodinated using a stoichiometric amount of nonradioactive cold NaI 
      to minimize multiple iodinations. Of the four tyrosines present in BDNF--Tyr-52, 
      Tyr-54, Tyr-63, and Tyr-86--only Tyr-63 and Tyr-86 were iodinated under the 
      experimental conditions used. Iodination of Tyr-63 resulted in modification 
      without alteration of the biological activity, whereas iodination of Tyr-86 
      resulted in a molecule with highly compromised biological activity. Similar 
      inactivation was observed if both Tyr-63 and Tyr-86 were iodinated. These 
      modified proteins exhibited conformation and dimerization apparently identical to 
      those of the native protein, as demonstrated by analytical ultracentrifugation, 
      gel filtration, light scattering, and circular dichroism. From these results, we 
      concluded that Tyr-52 and Tyr-54 are not accessible to the reagent and are 
      probably buried in the hydrophobic core, whereas Tyr-63 and Tyr-86 are exposed on 
      the surface of the molecule; of the two exposed residues, only Tyr-86 contributes 
      to the biological activity.
FAU - Rosenfeld, R
AU  - Rosenfeld R
AD  - Amgen Inc., Amgen Center, Thousand Oaks, California 91320-1789.
FAU - Philo, J S
AU  - Philo JS
FAU - Haniu, M
AU  - Haniu M
FAU - Stoney, K
AU  - Stoney K
FAU - Rohde, M F
AU  - Rohde MF
FAU - Wu, G M
AU  - Wu GM
FAU - Narhi, L O
AU  - Narhi LO
FAU - Wong, C
AU  - Wong C
FAU - Boone, T
AU  - Boone T
FAU - Hawkins, N N
AU  - Hawkins NN
AU  - et al.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Protein Sci
JT  - Protein science : a publication of the Protein Society
JID - 9211750
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 42HK56048U (Tyrosine)
RN  - 9679TC07X4 (Iodine)
RN  - EC 3.4.23.1 (Pepsin A)
RN  - F5WR8N145C (Sodium Iodide)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites
MH  - Brain-Derived Neurotrophic Factor
MH  - Chickens
MH  - Chromatography, High Pressure Liquid
MH  - Circular Dichroism
MH  - Ganglia/drug effects/ultrastructure
MH  - Humans
MH  - Iodine/*metabolism
MH  - Macromolecular Substances
MH  - Mass Spectrometry
MH  - Molecular Sequence Data
MH  - Nerve Tissue Proteins/chemistry/*metabolism/pharmacology
MH  - Pepsin A/metabolism
MH  - Peptide Mapping
MH  - Protein Conformation
MH  - Recombinant Proteins/metabolism
MH  - Sodium Iodide/metabolism
MH  - Structure-Activity Relationship
MH  - Tyrosine/metabolism
MH  - Ultracentrifugation
PMC - PMC2142266
EDAT- 1993/10/01 00:00
MHDA- 1993/10/01 00:01
CRDT- 1993/10/01 00:00
PHST- 1993/10/01 00:00 [pubmed]
PHST- 1993/10/01 00:01 [medline]
PHST- 1993/10/01 00:00 [entrez]
AID - 10.1002/pro.5560021012 [doi]
PST - ppublish
SO  - Protein Sci. 1993 Oct;2(10):1664-74. doi: 10.1002/pro.5560021012.