PMID- 9398330
OWN - NLM
STAT- MEDLINE
DCOM- 19980127
LR  - 20131121
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 36
IP  - 50
DP  - 1997 Dec 16
TI  - Localization of the heme binding region in soluble guanylate cyclase.
PG  - 15959-64
AB  - Soluble guanylate cyclase (sGC) is a heterodimeric hemoprotein composed of alpha1 
      and beta1 subunits. sGC is activated by nitric oxide (NO) and therefore plays a 
      central role in NO signal transduction. Activation of sGC by NO is believed to be 
      mediated by the interaction between NO and the heme of sGC. Spectroscopic and 
      kinetic studies have shown that the heme of sGC is in a unique environment. 
      Characterization of the heme environment is critical to the understanding of the 
      mechanism of NO activation. To approach this goal, the beta1 N-terminal fragment 
      consisting of residues 1-385 [beta1(1-385)] of sGC was expressed in E. coli. 
      beta1(1-385) was then purified to homogeneity in two steps by DEAE ion exchange 
      and gel filtration chromatography. Purified beta1(1-385) was found to contain a 
      stoichiometric amount of heme. The UV-visible spectrum of beta1(1-385) is almost 
      identical to that of the native heterodimeric sGC purified from bovine lung. 
      beta1(1-385) binds both NO and CO, leading to a shift in the Soret maximum from 
      431 nm to 398 and 423 nm, respectively. These spectral shifts are identical to 
      those observed with heterodimeric sGC purified from bovine lung. These results 
      suggest that the heme in the beta1(1-385) is similar to that in the heterodimeric 
      sGC. Therefore, for the first time, the heme binding region of sGC has been 
      unambiguously localized to the N-terminal region of the beta1 subunit. Our data 
      also suggest that the N-terminal region of the beta1 subunit of sGC is itself 
      sufficient for heme binding.
FAU - Zhao, Y
AU  - Zhao Y
AD  - Department of Biological Chemistry, University of Michigan Medical School, 
      University of Michigan, Ann Arbor, Michigan 48109-1065, USA.
FAU - Marletta, M A
AU  - Marletta MA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Hemeproteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 31C4KY9ESH (Nitric Oxide)
RN  - 42VZT0U6YR (Heme)
RN  - 7U1EE4V452 (Carbon Monoxide)
RN  - EC 4.6.1.2 (Guanylate Cyclase)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - Blotting, Western
MH  - Carbon Monoxide/metabolism
MH  - Cattle
MH  - Dimerization
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Escherichia coli/genetics
MH  - Guanylate Cyclase/*chemistry/metabolism
MH  - Heme/*chemistry/metabolism
MH  - Hemeproteins/chemistry
MH  - Lung/enzymology
MH  - Nitric Oxide/metabolism
MH  - Peptide Fragments/chemistry
MH  - Protein Binding
MH  - Rats
MH  - Recombinant Proteins/chemistry/isolation & purification
MH  - Solubility
MH  - Spectrophotometry
EDAT- 1998/01/31 00:00
MHDA- 1998/01/31 00:01
CRDT- 1998/01/31 00:00
PHST- 1998/01/31 00:00 [pubmed]
PHST- 1998/01/31 00:01 [medline]
PHST- 1998/01/31 00:00 [entrez]
AID - bi971825x [pii]
AID - 10.1021/bi971825x [doi]
PST - ppublish
SO  - Biochemistry. 1997 Dec 16;36(50):15959-64. doi: 10.1021/bi971825x.