Open AccessAnalysis of the aliphatic 1 H‐NMR spectrum of plasminogen kringle 4
Author(s) -
PETROS Andrew M.,
GYENES Marianne,
PATTHY László,
LLINÁS Miguel
Publication year - 1988
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1988.tb13734.x
Subject(s) - chemistry , kringle domain , proton nmr , two dimensional nuclear magnetic resonance spectroscopy , stereochemistry , nuclear overhauser effect , spectral line , lysine , chemical shift , nuclear magnetic resonance spectroscopy , physics , amino acid , biochemistry , gene , astronomy , recombinant dna
The aliphatic 1 H‐NMR spectrum of the kringle 4 domain of human plasminogen has been studied via two‐dimensional chemical shift correlated (COSY) and nuclear Overhauser correlated (NOESY) experiments at 300 MHz and 620 MHz. A number of aliphatic proton spin systems have been identified and several definite assignments have been made. This was mainly achieved by comparison of the human kringle 4 spectrum with spectra of the porcine, bovine and chicken homologs and also with that of the kringle 1 from human plasminogen on which we have reported previously. The three valyl and two leucyl residues of human kringle 4 have been assigned. The eleven threonyl spin systems have been identified via a RELAYED‐COSY experiment and Thr 17 has been assigned. The three alanyl spin systems have been identified and assigned. Six seryl spin systems have been identified and the signals from the seven glycyl residues of human kringle 4 have been located with Gly 45 assigned. Furthermore, 24 AMX spin systems have been mapped in the COSY spectrum of human kringle 4 and H α ‐H β,β′ spin systems of Tyr 2 , Tyr 41 , Tyr 50 , Tyr 74 , Trp 25 and Trp 62 have been assigned. From the spectrum of a deglycosylated chicken homolog, the ɛ‐methyl singlets of Met 28 and Met 48 have been assigned. Finally, ligand effects on selected aliphatic resonances were observed which could be analyzed in terms of residues likely to neighbor the kringle lysine‐binding site.