Catecholamine-containing proteins from the pharate pupal cuticle of the tobacco hornworm, Manduca sexta.
Three catecholamine-containing proteins from tanning, pharate pupal, abdominal cuticle of the tobacco hornworm, Manduca sexta, have been purified to apparent homogeneity and characterized. These proteins have apparent molecular masses of 32, 41 and 48 kDa and were shown by liquid chromatography-electrochemical analysis, after heating in 1 M acetic acid at 110 degrees C, to release nu-beta-alanylnorepinephrine (NBANE). NBANE is a hydrolysis product of N-beta-alanyldopamine (NBAD) that is bonded covalently at the beta-side-chain carbon to amino acid residues of cuticular proteins. Amino acid compositional analysis revealed that MS-PCP32 (32 KDa) has a high content of alanine (25.6%), valine (13.1%), proline (10.8%) and glycine (10%), a low level of phenylalanine, and no detectable tyrosine or methionine. In contrast, MS-PCP41 (41 kDa) had a much higher content of glycine (31.2%) and substantial levels of serine (9.2%), proline (10.2%) and glutamate/glutamine (10.6%), whereas tyrosine and phenylalanine were not detected. Two of the three purified proteins showed apparent similarities to each other in nu-terminal amino acid sequences, and to several other known insect cuticular proteins. Proteins MS-PCP41 and MS-PCP48 had the characteristic GGX triplet repeat, which is found in a variety of cuticular proteins and may be important for protein folding appropriate for cuticular functions. Therefore, a diversity of cuticular proteins with different amino acid sequences and properties apparently are secreted into the presumptive pupal exocuticle. These then can form adducts and possible cross-links with NBAD through its quinonoid intermediates during cuticular sclerotization.
Okot-Kotber
B M
BM
Department of Entomology, Kansas State University, Manhattan 66506, USA.
Morgan
T D
TD
Hopkins
T L
TL
Kramer
K J
KJ
D000595:Amino Acid Sequence; D000818:Animals; D002395:Catecholamines / Q000032:analysis*; D018402:Manduca / Q000737:chemistry*; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D011679:Pupa
D002395:Catecholamines; D011506:Proteins