After a beetle first molts, its exoskeleton is soft and hydrated. Somehow, it dries out and forms a hard, stiff exoskeleton. Since the 1940s, scientists have wondered which enzyme among several possible candidates was involved in the hardening process.
The K-State researchers have found that by knocking out an enzyme called laccase-2, cuticle tanning, the process of hardening and pigmentation, can be prevented in the red flour beetle, Tribolium castaneum.
A paper, to be released this week in the Proceedings of the National Academy of Sciences, presents the research results. The K-State researchers are Yasuyuki Arakane, research associate in biochemistry; Subbaratnam Muthukrishnan, professor of biochemistry; Richard Beeman, adjunct professor of entomology; Michael Kanost, professor and head of the department of biochemistry; and Karl Kramer, adjunct professor emeritus of biochemistry.
Kramer said K-State researchers wanted to find out what happens between the times when the cuticle is soft and when it is hard. They studied the cuticle's composition and how the components interacted to give it stiffness, flexibility and lightness. The main components in the cuticle are proteins and chitin, which also are found in crustaceans and other invertebrates.
The researchers knew one of two classes of oxidative enzymes, tyrosinases or laccases, is likely responsible for catalyzing the exoskeleton's hardening by cross-linking cuticular proteins, Kanost said.
"When we knocked out tyrosinase, everything was normal," Kramer said. "When we knocked out laccase-2, we prevented tanning from taking place."
When the laccase-2 gene was not expressed, the newly formed cut
Source:Kansas State University