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Lima bean proteinase inhibitor. Origins of circular dichroism bands and modification by Br/sub 2//sup -/ and (CNS)/sub 2//sup -/

Journal Article · · J. Biol. Chem.; (United States)
OSTI ID:5376549

Origins of CD bands in lima bean proteinase inhibitor were deduced from an acetylation-deacetylation study of the sole tyrosyl residue in the protein (Tyr 69), and by analogy with Bowman-Birk soybean proteinase inhibitor. A broad negative CD band centered around 280 nm arises mainly from the longest wavelength transition of cystinyl side chains. The second cystinyl transition gives rise to a positive CD band of a comparable intensity at 247 nm. the /sup 1/L/sub b/ vibronic transition of Tyr 69 has negative CD around 280 nm, contributing approximately 10% of the total CD intensity at 278 nm. the 232 nm positive shoulder is from the /sup 1/L/sub a/ vibronic transition of Tyr 69. Radical anions, Br/sub 2//sup -/ and (CNS)/sub 2/, generated by the irradiation of N/sub 2/O-saturated inhibitor solutions containing KBr or KCNS, reduced tyrosyl CD without affecting disulfide CD bands, indicating that the radical anions damaged Tyr 69 without altering protein conformation. The inhibitor modified at Tyr 69 by Br/sub 2//sup -/ and (CNS)/sub 2//sup -/ retained full activity toward trypsin and chymotrypsin. The irradiation of the inhibitor in the air saturated solution led to loss in tyrosyl as well as cystinl CD bands and decline in both antiproteinase activities.

Research Organization:
Univ. of California, Los Angeles
DOE Contract Number:
EY-76-C-03-0012
OSTI ID:
5376549
Journal Information:
J. Biol. Chem.; (United States), Journal Name: J. Biol. Chem.; (United States) Vol. 254:16; ISSN JBCHA
Country of Publication:
United States
Language:
English