Bleaching kinetics of artificial visual pigments with modifications near the ring-polyene chain connection.
Absorbance difference spectra were recorded at 20 degrees C from 30 ns to milliseconds after photolysis of lauryl maltoside suspensions of artificial visual pigments derived from 9-cis isomers of 5-ethylretinal, 8,16-methanoretinal (a 6-s-trans-bicyclic analogue), or 5-demethyl-8-methylretinal. In all three pigments, the earliest intermediate that was detected had the characteristics of a mixture of bathorhodopsin and a blue-shifted intermediate, BSI, which is the first decay product of bathorhodopsin in bovine rhodopsin. The first decays resolved on the nanosecond time scale were the formation of the lumirhodopsin analogues. Subsequent decays were able to be fit with a mechanistic scheme which has been shown to apply to both membrane and detergent suspensions of rhodopsin. Large increases were seen in the amount of metarhodopsin I which appeared after photolysis of 5-ethylisorhodopsin and the bicyclic isorhodopsin analogue, while 5-demethyl-8-methylisorhodopsin more closely followed native rhodopsin in decaying through meta I380, a 380 nm absorbing precursor to metarhodopsin II. In addition to forming more metarhodopsin I, the bicyclic analogue stabilized the metarhodopsin I-metarhodopsin II equilibrium similarly to what has been previously reported for 9-demethylrhodopsin in detergent, introducing the possibility that the bicyclic analogue could similarly be defective in transducin activation. These observations support the idea that long after initial photolysis, structural details of the retinylidene chromophore continue to play a decisive role in processes leading to the activated form, metarhodopsin II.
Szundi
Istvan
I
Department of Chemistry and Biochemistry, University of California, Santa Cruz, California 95064, USA.
de Lera
Angel R
AR
Pazos
Yolanda
Y
Alvarez
Rosana
R
Oliana
Marco
M
Sheves
Mordechai
M
Lewis
James W
JW
Kliger
David S
DS
D000818:Animals; D002417:Cattle; D066298:In Vitro Techniques; D007700:Kinetics; D010777:Photochemistry; D010782:Photolysis; D012168:Retinal Pigments / Q000138:chemical synthesis / Q000737:chemistry* / Q000528:radiation effects*; D012172:Retinaldehyde / Q000031:analogs & derivatives / Q000737:chemistry / Q000528:radiation effects; D012243:Rhodopsin / Q000031:analogs & derivatives / Q000737:chemistry / Q000528:radiation effects; D013053:Spectrophotometry
D012168:Retinal Pigments; C042882:isorhodopsin; C013110:metarhodopsins; D012243:Rhodopsin; D012172:Retinaldehyde