TY - JOUR
T1 - Unraveling the sequence and structure of the protein osteocalcin from a 42 ka fossil horse
AU - Ostrom, P H
AU - Gandhi, H
AU - Strahler, J R
AU - Walker, A K
AU - Andrews, P C
AU - Leykam, J
AU - Stafford, T W
AU - Kelly, R L
AU - Walker, D N
AU - Buckley, M
AU - Humpula, J
PY - 2006/4/15
Y1 - 2006/4/15
N2 - We report the first complete amino acid sequence and evidence of secondary structure for osteocalcin from a temperate fossil. The osteocalcin derives from a 42 ka equid bone excavated from Juniper Cave, Wyoming. Results were determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-MS) and Edman sequencing with independent confirmation of the sequence in two laboratories. The ancient sequence was compared to that of three modern taxa: horse (Eguus caballtts), zebra (Eguus grevyi), and donkey (Eduus asinus). Although there was no difference in sequence among modern taxa, MALDI-MS and Edman sequencing show that residues 48 and 49 of our modern horse are Thr, Ala rather than Pro, Val as previously reported (Carstanjen B., Wattiez, R., Armory, H., Lepage, O.M., Remy, B., 2002. Isolation and characterization of equine osteocalcin. Ann. Med. Vet. 146(1), 31-38). MALDI-MS and Edman sequencing data indicate that the osteocalcin sequence of the 42 ka fossil is similar to that of modern horse. Previously inaccessible structural attributes for ancient osteocalcin were observed. Glu(39) rather than Gln(39) is consistent with deamidation, a process known to occur during fossilization and aging. Two post-translational modifications were documented: Hyp(9) and a disulfide bridge. The latter suggests at least partial retention of secondary structure. As has been done for ancient DNA research, we recommend standards for preparation and criteria for authenticating results of ancient protein sequencing. (c) 2006 Elsevier Inc. All rights reserved.
AB - We report the first complete amino acid sequence and evidence of secondary structure for osteocalcin from a temperate fossil. The osteocalcin derives from a 42 ka equid bone excavated from Juniper Cave, Wyoming. Results were determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-MS) and Edman sequencing with independent confirmation of the sequence in two laboratories. The ancient sequence was compared to that of three modern taxa: horse (Eguus caballtts), zebra (Eguus grevyi), and donkey (Eduus asinus). Although there was no difference in sequence among modern taxa, MALDI-MS and Edman sequencing show that residues 48 and 49 of our modern horse are Thr, Ala rather than Pro, Val as previously reported (Carstanjen B., Wattiez, R., Armory, H., Lepage, O.M., Remy, B., 2002. Isolation and characterization of equine osteocalcin. Ann. Med. Vet. 146(1), 31-38). MALDI-MS and Edman sequencing data indicate that the osteocalcin sequence of the 42 ka fossil is similar to that of modern horse. Previously inaccessible structural attributes for ancient osteocalcin were observed. Glu(39) rather than Gln(39) is consistent with deamidation, a process known to occur during fossilization and aging. Two post-translational modifications were documented: Hyp(9) and a disulfide bridge. The latter suggests at least partial retention of secondary structure. As has been done for ancient DNA research, we recommend standards for preparation and criteria for authenticating results of ancient protein sequencing. (c) 2006 Elsevier Inc. All rights reserved.
KW - ANCIENT DNA
KW - BONE
KW - RECOGNITION
U2 - 10.1016/j.gca.2006.01.004
DO - 10.1016/j.gca.2006.01.004
M3 - Article
SN - 0016-7037
VL - 70
SP - 2034
EP - 2044
JO - Geochimica et Cosmochimica Acta
JF - Geochimica et Cosmochimica Acta
IS - 8
ER -