PMID- 3768308
OWN - NLM
STAT- MEDLINE
DCOM- 19861124
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 25
IP  - 17
DP  - 1986 Aug 26
TI  - Photoaffinity labeling of the rat plasma vitamin D binding protein with 
      [26,27-3H]-25-hydroxyvitamin D3 3 beta-[N-(4-azido-2-nitrophenyl)glycinate].
PG  - 4729-33
AB  - It is well recognized that the vitamin D binding protein (DBP) is important for 
      the transport of vitamin D, 25-hydroxyvitamin D (25-OH-D), and its metabolites. 
      In an attempt to better understand the molecular-binding properties of this 
      ubiquitous protein, we designed and synthesized a photoaffinity analogue of 
      25-OH-D3 and its radiolabeled counterpart. This analogue, 25-hydroxyvitamin D3 3 
      beta-[N-(4-azido-2-nitrophenyl)glycinate] (25-OH-D3-ANG), was recognized by the 
      rat DBP and was about 10 times less active than 25-OH-D3 in terms of binding. 
      Incubation of [3H]25-OH-D3 or [3H]25-OH-D3-ANG with rat DBP revealed that both 
      compounds were specifically bound to a protein with a sedimentation coefficient 
      of 4.1 S. Each was displaced with a 500-fold excess of 25-OH-D3. When 
      [3H]25-OH-D3-ANG was exposed to UV radiation in the presence of rat DBP followed 
      by the addition of a 500-fold excess of 25-OH-D3, there was no displacement of 
      tritium from the 4.1S peak. Sodium dodecyl sulfate-polyacrylamide gel 
      electrophoresis autoradiographic analysis of [3H]25-OH-D3-ANG exposed to UV 
      radiation in the presence of rat DBP followed by the addition of a 500-fold 
      excess of 25-OH-D3 revealed one major band with a molecular weight of 52 000. 
      These data provide strong evidence that [3H]25-OH-D3-ANG was covalently linked to 
      the rat DBP. This photoaffinity probe should provide a valuable tool for the 
      analysis of the binding site on this transport protein.
FAU - Ray, R
AU  - Ray R
FAU - Holick, S A
AU  - Holick SA
FAU - Hanafin, N
AU  - Hanafin N
FAU - Holick, M F
AU  - Holick MF
LA  - eng
GR  - AM36963/AM/NIADDK NIH HHS/United States
GR  - AM36985/AM/NIADDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Affinity Labels)
RN  - 0 (Azides)
RN  - 0 (Carbon Radioisotopes)
RN  - 0 (Indicators and Reagents)
RN  - 0 (Vitamin D-Binding Protein)
RN  - 10028-17-8 (Tritium)
RN  - 101396-04-7 (25-hydroxyvitamin D3 3-(N-(4-azido-2-nitrophenyl)glycinate))
RN  - P6YZ13C99Q (Calcifediol)
SB  - IM
MH  - Affinity Labels/*chemical synthesis
MH  - Animals
MH  - Azides/*chemical synthesis/metabolism
MH  - Binding, Competitive
MH  - Calcifediol/*analogs & derivatives/chemical synthesis/metabolism
MH  - Carbon Radioisotopes
MH  - Indicators and Reagents
MH  - Kinetics
MH  - Rats
MH  - Tritium
MH  - Vitamin D-Binding Protein/*blood/isolation & purification
EDAT- 1986/08/26 00:00
MHDA- 1986/08/26 00:01
CRDT- 1986/08/26 00:00
PHST- 1986/08/26 00:00 [pubmed]
PHST- 1986/08/26 00:01 [medline]
PHST- 1986/08/26 00:00 [entrez]
AID - 10.1021/bi00365a001 [doi]
PST - ppublish
SO  - Biochemistry. 1986 Aug 26;25(17):4729-33. doi: 10.1021/bi00365a001.