PMID- 18547058
OWN - NLM
STAT- MEDLINE
DCOM- 20080716
LR  - 20231213
IS  - 1520-4995 (Electronic)
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 47
IP  - 27
DP  - 2008 Jul 8
TI  - Probing the steroid binding domain-like I (SBDLI) of the sigma-1 receptor binding 
      site using N-substituted photoaffinity labels.
PG  - 7205-17
LID - 10.1021/bi800564j [doi]
AB  - Radioiodinated photoactivatable photoprobes can provide valuable insights 
      regarding protein structure. Previous work in our laboratory showed that the 
      cocaine derivative and photoprobe 3-[ (125)I]iodo-4-azidococaine ([ (125)I]IACoc) 
      binds to the sigma-1 receptor with 2-3 orders of magnitude higher affinity than 
      cocaine [Kahoun, J. R. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 1393-1397]. Using 
      this photoprobe, we demonstrated the insertion site for [ (125)I]IACoc to be 
      Asp188 [Chen, Y. (2007) Biochemistry 46, 3532-3542], which resides in the 
      proposed steroid binding domain-like II (SBDLII) region (amino acids 176-194) 
      [Pal, A. (2007) Mol. Pharmacol. 72, 921-933]. An additional photoprobe based on 
      the sigma-1 receptor ligand fenpropimorph, 1- N-(2-3-[ (125)I]iodophenyl)propane 
      ([ (125)I]IAF), was found to label a peptide in both the SBDLII and steroid 
      binding domain-like I (SBDLI) (amino acids 91-109) [Pal, A. (2007) Mol. 
      Pharmacol. 72, 921-933]. In this report, we describe two novel strategically 
      positioned carrier-free, radioiodinated photoaffinity labels specifically 
      designed to probe the putative "nitrogen interacting region" of sigma-1 receptor 
      ligands. These two novel photoprobes are (-)-methyl 
      3-(benzoyloxy)-8-2-(4-azido-3-[ 
      (125)I]iodobenzene)-1-ethyl-8-azabicyclo[3.2.1]octane-2-carboxylate ([ 
      (125)I]-N-IACoc) and N-propyl- 
      N-(4-azido-3-iodophenylethyl)-3-(4-fluorophenyl)propylamine ([ (125)I]IAC44). In 
      addition to reporting their binding affinities to the sigma-1 and sigma-2 
      receptors, we show that both photoaffinity labels specifically and covalently 
      derivatized the pure guinea pig sigma-1 receptor (26.1 kDa) [Ramachandran, S. 
      (2007) Protein Expression Purif. 51, 283-292]. Cleavage of the photolabeled 
      sigma-1 receptor using Endo Lys C and cyanogen bromide (CNBr) revealed that the [ 
      (125)I]-N-IACoc label was located primarily in the N-terminus and 
      SBDLI-containing peptides of the sigma-1 receptor, while [ (125)I]IAC44 was found 
      in peptide fragments consistent with labeling of both SBDLI and SBDLII.
FAU - Fontanilla, Dominique
AU  - Fontanilla D
AD  - Department of Pharmacology, University of Wisconsin School of Medicine and Public 
      Health, Madison, Wisconsin 53706, USA.
FAU - Hajipour, Abdol R
AU  - Hajipour AR
FAU - Pal, Arindam
AU  - Pal A
FAU - Chu, Uyen B
AU  - Chu UB
FAU - Arbabian, Marty
AU  - Arbabian M
FAU - Ruoho, Arnold E
AU  - Ruoho AE
LA  - eng
GR  - T32 GM008688/GM/NIGMS NIH HHS/United States
GR  - F31 DA022932/DA/NIDA NIH HHS/United States
GR  - F31 DA022932-02/DA/NIDA NIH HHS/United States
GR  - R01 MH065503-04/MH/NIMH NIH HHS/United States
GR  - T32 GM08688/GM/NIGMS NIH HHS/United States
GR  - R01 MH065503-05/MH/NIMH NIH HHS/United States
GR  - T32 GM008688-15/GM/NIGMS NIH HHS/United States
GR  - F31 DA022932-03/DA/NIDA NIH HHS/United States
GR  - T32 GM008688-14/GM/NIGMS NIH HHS/United States
GR  - R01 MH065503/MH/NIMH NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20080612
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Peptides)
RN  - 0 (Photoaffinity Labels)
RN  - 0 (Receptors, sigma)
RN  - 0 (sigma-2 receptor)
RN  - EC 3.4.24.- (Metalloendopeptidases)
RN  - EC 3.4.24.20 (peptidyl-Lys metalloendopeptidase)
RN  - I5Y540LHVR (Cocaine)
RN  - OS382OHJ8P (Cyanogen Bromide)
SB  - IM
MH  - Animals
MH  - Autoradiography
MH  - Binding Sites
MH  - Cocaine/analogs & derivatives/chemical synthesis/chemistry
MH  - Cyanogen Bromide/metabolism
MH  - Guinea Pigs
MH  - Metalloendopeptidases/metabolism
MH  - Molecular Weight
MH  - Peptides/metabolism
MH  - Photoaffinity Labels/*metabolism
MH  - Protein Structure, Tertiary
MH  - Rats
MH  - Receptors, sigma/*chemistry/metabolism
MH  - Sigma-1 Receptor
PMC - PMC3250216
MID - NIHMS335124
EDAT- 2008/06/13 09:00
MHDA- 2008/07/17 09:00
PMCR- 2012/01/03
CRDT- 2008/06/13 09:00
PHST- 2008/06/13 09:00 [pubmed]
PHST- 2008/07/17 09:00 [medline]
PHST- 2008/06/13 09:00 [entrez]
PHST- 2012/01/03 00:00 [pmc-release]
AID - 10.1021/bi800564j [doi]
PST - ppublish
SO  - Biochemistry. 2008 Jul 8;47(27):7205-17. doi: 10.1021/bi800564j. Epub 2008 Jun 
      12.