PMID- 10608872
OWN - NLM
STAT- MEDLINE
DCOM- 20000208
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 53
DP  - 1999 Dec 31
TI  - Definition of the interaction domain for cytochrome c on cytochrome c oxidase. I. 
      Biochemical, spectral, and kinetic characterization of surface mutants in subunit 
      ii of Rhodobacter sphaeroides cytochrome aa(3).
PG  - 38032-41
AB  - To determine the interaction site for cytochrome c (Cc) on cytochrome c oxidase 
      (CcO), a number of conserved carboxyl residues in subunit II of Rhodobacter 
      sphaeroides CcO were mutated to neutral forms. A highly conserved tryptophan, 
      Trp(143), was also mutated to phenylalanine and alanine. Spectroscopic and metal 
      analyses of the surface carboxyl mutants revealed no overall structural changes. 
      The double mutants D188Q/E189N and D151Q/E152N exhibit similar steady-state 
      kinetic behavior as wild-type oxidase with horse Cc and R. sphaeroides Cc(2), 
      showing that these residues are not involved in Cc binding. The single mutants 
      E148Q, E157Q, D195N, and D214N have decreased activities and increased K(m) 
      values, indicating they contribute to the Cc:CcO interface. However, their 
      reactions with horse and R. sphaeroides Cc are different, as expected from the 
      different distribution of surface lysines on these cytochromes c. Mutations at 
      Trp(143) severely inhibit activity without changing the K(m) for Cc or disturbing 
      the adjacent Cu(A) center. From these data, we identify a Cc binding area on CcO 
      with Trp(143) and Asp(214) close to the site of electron transfer and Glu(148), 
      Glu(157), and Asp(195) providing electrostatic guidance. The results are 
      completely consistent with time-resolved kinetic measurements (Wang, K., Zhen, 
      Y., Sadoski, R., Grinnell, S., Geren, L., Ferguson-Miller, S., Durham, B., and 
      Millett, F. (1999) J. Biol. Chem. 274, 38042-38050) and computational docking 
      analysis (Roberts, V. A., and Pique, M. E. (1999) J. Biol. Chem. 274, 
      38051-38060).
FAU - Zhen, Y
AU  - Zhen Y
AD  - Department of Biochemistry, Michigan State University, East Lansing, Michigan 
      48824, USA.
FAU - Hoganson, C W
AU  - Hoganson CW
FAU - Babcock, G T
AU  - Babcock GT
FAU - Ferguson-Miller, S
AU  - Ferguson-Miller S
LA  - eng
GR  - GM25480/GM/NIGMS NIH HHS/United States
GR  - GM26916/GM/NIGMS NIH HHS/United States
GR  - GM57323/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Cytochrome c Group)
RN  - EC 1.9.3.1 (Electron Transport Complex IV)
SB  - IM
MH  - Animals
MH  - Cytochrome c Group/*metabolism
MH  - Electron Spin Resonance Spectroscopy
MH  - Electron Transport
MH  - Electron Transport Complex IV/genetics/isolation & purification/*metabolism
MH  - Horses
MH  - Kinetics
MH  - Models, Molecular
MH  - Osmolar Concentration
MH  - Rhodobacter sphaeroides/*enzymology
MH  - Spectroscopy, Near-Infrared
EDAT- 1999/12/23 00:00
MHDA- 1999/12/23 00:01
CRDT- 1999/12/23 00:00
PHST- 1999/12/23 00:00 [pubmed]
PHST- 1999/12/23 00:01 [medline]
PHST- 1999/12/23 00:00 [entrez]
AID - S0021-9258(19)52992-1 [pii]
AID - 10.1074/jbc.274.53.38032 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Dec 31;274(53):38032-41. doi: 10.1074/jbc.274.53.38032.