The stable carotenoid cation radical (Car(.+)) and chlorophyll cation radic
al (Chl(z)(.+)) in photosystem II (PS II) have been studied by pulsed elect
ron nuclear double resonance (ENDOR) spectroscopy. The spectra were essenti
ally the same for oxygen-evolving PS II and Mn-depleted PS II. The radicals
were generated by illumination given at low temperatures, and the ENDOR sp
ectra were attributed to Car(.+) and Chl(z)(.+) on the basis of their chara
cteristic behavior with temperature as demonstrated earlier [Hanley et al.
(1999) Biochemistry 38, 8189-8195]: i.e., (a) the Car(.+) alone was generat
ed by illumination at less than or equal to 20 K, while Chl(z)(.+) alone wa
s generated at 200 K, and (b) warming of the sample containing the Car(.+)
to 200 K resulted in the loss of the signal attributable to Car(.+) and its
replacement by a spectrum attributable to the Chl(z)(.+). A map of the hyp
erfine structure of Car(.+) in PS II and in organic solvent was obtained. T
he largest observed hyperfine splitting for Car(.+) in either environment w
as in the order of 8-9 MHz. Thus, the spin density on the cation is propose
d to be delocalized over the carotenoid molecule. The pulsed ENDOR spectrum
of Chl(z)(.+) was compared to that obtained from a Chl a cation in frozen
organic solvent. The hyperfine coupling constants attributed to the beta -p
rotons at position 17 and 18 are well resolved from Chl(z)(.+) in PS II (10
.8 and 14.9 MHz) but not in Chl a(.+) in organic solvent (12.5 MHz). This s
uggests a more defined conformation of ring IV with respect to the rest of
the tetrapyrrole ring plane of Chl(z)(.+) than Chl a(.+) probably induced b
y the protein matrix.