The mass spectrometric analysis of the major chain d1, one of the four heme-binding chains in earthworm hemeglobin of Lumbricus terrestris, revealed a difference between the measured and the sequence derived masses indicating errors in the reported sequence. The purpose of this study is to clarify the possible sequence ambiguity by mass spectrometric peptide mapping experiments. After the digestion of chain d with a number of enzymes, proteolytic peptides in the region of 51-140 were identified and indicated a correct sequence assignment in this region. In addition, digestion with V8 protease revealed an sequence error in position 58 (E instead of Q). N-terminal Edman sequencing allowed the correction of misidentified amino acids 7 (G7S), 23 (Q23E) and 33 (K33R). The refined sequence (1-140) results in a molecular mass of 15961 Da which is 28 Da below the measured mass of 15989Da. A paper describing the resolution of these discrepancies has been published Q. Xie, R. A. Donahue, Jr., K. Schneider, U.A. Mirza, I. Haller, B.T. Chait & A. F. Riggs """"""""Structure of chain d of the gigantic hemoglobin of the earthworm"""""""" BBA 1337 (1997) 241-247.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR000862-27
Application #
6307605
Study Section
Project Start
1999-12-01
Project End
2000-11-30
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
27
Fiscal Year
2000
Total Cost
$8,199
Indirect Cost
Name
Rockefeller University
Department
Type
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
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