Eggshell precursor proteins of Fasciola hepatica, II. Microheterogeneity in vitelline protein B.
At least 3 structural protein precursors of the eggshell are synthesized and stockpiled in the extensive vitelline cells of the liver fluke Fasciola hepatica L. One of these, vitelline protein B, consists of a closely related family of proteins that owes its apparent electrophoretic heterogeneity to variations in the Tyr to DOPA conversion as well as to subtle variations in the primary sequence. The efficiency of the Tyr to DOPA conversion ranges from a maximum of about 90% to a minimum of 55% in the protein. Trypsin digestion in borate buffer at pH 8 was used to produce DOPA-peptides for sequencing. Notably, trypsin does not cleave Arg/Lys-DOPA sequences at borate concentrations greater than 0.15 M. Peptides with DOPA-containing sequences most frequently have flanking amino acids such as Lys, Ser, or Asp on the N-terminal side and Gly or Asp on the C-terminal side. All protein variants fall within a narrow molecular weight range (30-33 kDa), a pI range of 6.9 to 8.3, and the collective majority would appear to share a common N-terminal sequence up to residue 28. The results suggest some combination of the following: variations in post-translational hydroxylation, alternative post-transcriptional splicing and/or the existence of multiple gene copies of eggshell precursors. The latter have been shown to occur in the blood fluke Schistosoma mansoni [15].
Waite
J H
JH
Marine Biology/Biochemistry Program, College of Marine Studies, University of Delaware, Lewes 19958.
Rice-Ficht
A C
AC
D000595:Amino Acid Sequence; D000818:Animals; D004295:Dihydroxyphenylalanine / Q000032:analysis; D004527:Egg Proteins / Q000737:chemistry* / Q000302:isolation & purification; D005210:Fasciola hepatica / Q000737:chemistry*; D015801:Helminth Proteins / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010063:Ovum / Q000737:chemistry; D014357:Trypsin; D014817:Vitelline Membrane / Q000737:chemistry*
D004527:Egg Proteins; D015801:Helminth Proteins; C077550:vitelline protein B1, Fasciola hepatica; C077551:vitelline protein B2, Fasciola hepatica; D004295:Dihydroxyphenylalanine; D014357:Trypsin