Contribution of tryptophan residues to the combining site of a monoclonal anti dinitrophenyl spin-label antibody.

Content Provider	Semantic Scholar
Author	Anglister, Jacob A. Bond, Martha W. Frey, Tom Leahy, Denis Levitt, M. Mcconnell, H. M. Rule, Gordon S. Tomasello, J. Whittaker, Mark
Copyright Year	1987
Abstract	Two Fab fragments of the monoclonal anti dinitrophenyl (DNP) spin-label antibody AN02 were prepared by recombination of specifically deuterated heavy and light chains. In the recombinant H(I)L(II) all the tyrosines and phenylalanines were perdeuterated as were the tryptophan residues of the heavy chain. In the recombinant H(II)L(I) all the tyrosines and phenylalanines were perdeuterated as were the tryptophan residues of the light chain. Saturation of three resonances of H(I)L(II), assigned to tryptophan protons of the light chain, resulted in magnetization transfer to the aromatic proton at position 6 of the DNP ring and to the CH2 protons of the glycines linked to the DNP in a diamagnetic hapten (DNP-DG). Saturation of three resonances of H(II)L(I) assigned to tryptophan protons of the heavy chain resulted in magnetization transfer to the CH2 protons of the glycines in DNP-DG. From the dependence of the magnetization transfer on the irradiation time, the cross relaxation rates between the involved protons were estimated. The inferred distances between these protons of the hapten and certain tryptophan protons are 3-4 A. It is concluded that in the combining site of AN02 there is one tryptophan from the light chain and one tryptophan from the heavy chain that are very near the hapten. When all tyrosines and phenylalanines were perdeuterated and all tryptophan aromatic protons were deuterated except for the protons at positions 2 and 5, titration of the Fab fragments with variable amounts of paramagnetic hapten showed that one proton from the light chain tryptophan is near (less than 7 A) the unpaired electron and that three other protons are significantly closer than 15 A.(ABSTRACT TRUNCATED AT 250 WORDS)
Starting Page	75
Ending Page	86
Page Count	12
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://www.dtic.mil/get-tr-doc/pdf?AD=ADA219760&Location=U2&doc=GetTRDoc.pdf
Alternate Webpage(s)	http://www.dtic.mil/dtic/tr/fulltext/u2/a219760.pdf
PubMed reference number	3120771v1
Volume Number	26
Issue Number	19
Journal	Biochemistry
Language	English
Access Restriction	Open
Subject Keyword	2,4-Dinitrophenol AROMATICS Acetaminophen 32 MG/ML Oral Suspension Binding Sites CH2 Distance Inference PersonNameUse - assigned Protons Recombinants SLC3A2 gene Single-Chain Antibodies Titration Method Tryptophan decigram dinitrophenyl
Content Type	Text
Resource Type	Article