The proteolytic digestion of ox neurofilaments with trypsin and alpha-chymotrypsin.
Brief digestion of ox neurofilaments with trypsin liberates fragments that are soluble and have molecular weights ranging from 164 000 to 97 000. Peptide fingerprinting indicates that these regions, termed the tryptic head-regions, arise from the 205 000- and 158 000-mol.wt. components of the triplet. The remains of the parent polypeptides sediment with normal filaments and have been termed tail-regions. Digestion of neurofilaments with chymotrypsin also liberates soluble fragments (chymotryptic head-regions) but these have mol.wts. 171 000 and 119 000, though they too originate from the higher-molecular-weight triplet polypeptides. Tryptic and chymotryptic head-regions have extensive homology, and a low (less than or equal to 20%) helix content. Electron microscopy shows that chymotryptic digestion rapidly reduces the length of filaments, probably because this enzyme preferentially attacks the 72 000-mol.wt. polypeptide. In contrast, brief digestion with trypsin does not reduce filament length even though more than 90% of the two higher-molecular-weight components have been cleaved. These results indicate that the backbone of native filaments is formed from the 72 000-mol.wt. polypeptide together with the tail-regions from the 205 000- and 158 000-mol.wt. polypeptides. The corresponding head-regions of these components, which can represent nearly 75% of each molecule, are not necessary for preserving the backbone of native neurofilaments and are therefore good candidates for being the side arms that connect these filaments in nerve cells.
D000818:Animals; D002417:Cattle; D002918:Chymotrypsin / Q000494:pharmacology; D003599:Cytoskeleton / Q000032:analysis / Q000187:drug effects* / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D007381:Intermediate Filament Proteins; D008297:Male; D008854:Microscopy, Electron; D008970:Molecular Weight; D016900:Neurofilament Proteins; D010446:Peptide Fragments / Q000032:analysis; D011487:Protein Conformation; D013127:Spinal Nerves / Q000032:analysis / Q000648:ultrastructure; D014357:Trypsin / Q000494:pharmacology
D007381:Intermediate Filament Proteins; D016900:Neurofilament Proteins; D010446:Peptide Fragments; D002918:Chymotrypsin; D014357:Trypsin