The pancornulins: a group of small proline rich-related cornified envelope precursors with bifunctional capabilities in isopeptide bond formation.
In this report, the pancornulins are identified as members of the spr (small, proline-rich) multigene family by amino acid sequence and mass spectrometry analyses. One of the pancornulins (14.9 kDa) is identical to the protein predicted by spr-1 clone 128. The other pancornulins (16.9 kDa and 22 kDa) are novel members of the spr family. Immunoelectron microscopy of purified cornified envelopes with a pancornulin-specific antibody established these proteins more definitively as cornified envelope precursors. In addition, two-dimensional electrophoretic analyses of keratinocyte extracts labeled enzymatically with dansylcadaverine (to identify amine acceptors) or dansylPGGQQIV (to identify amine donors) showed that both glutamine and lysine residues within the pancornulins participate in the isopeptide linkage characteristic of cornified envelope formation. These results contrasted with those obtained using involucrin, a prominent cornified envelope protein shown capable of acting only as an amine acceptor in this system. Novel partial cDNAs obtained after reverse transcription and polymerase chain reaction amplification of total messenger RNA with pancornulin-specific primers suggest that the spr multigene family may be even larger than previously described. The bifunctional reactivity of the pancornulins in cross-linking and the large number of family members identified to date suggest that the pancornulins and other spr-1-related proteins may be more important in cornified envelope formation than previously considered, perhaps functioning as "bridge" molecules during the early phases of cornified envelope assembly.
Greco
M A
MA
Department of Dermatology, Harvard Medical School, Massachusetts General Hospital, Boston.
Lorand
L
L
Lane
W S
WS
Baden
H P
HP
Parameswaran
K N
KN
Kvedar
J C
JC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D055233:Cornified Envelope Proline-Rich Proteins; D018076:DNA, Complementary / Q000032:analysis; D004817:Epidermis / Q000737:chemistry*; D006801:Humans; D007525:Isoelectric Focusing; D015603:Keratinocytes; D008565:Membrane Proteins; D051379:Mice; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000737:chemistry; D011506:Proteins / Q000235:genetics; D017421:Sequence Analysis; D014466:Ultraviolet Rays
D055233:Cornified Envelope Proline-Rich Proteins; D018076:DNA, Complementary; D008565:Membrane Proteins; D011498:Protein Precursors; D011506:Proteins; C032253:involucrin