Amide-proton exchange of water-soluble proteins of different structural classes studied at the submolecular level by infrared spectroscopy.
For eleven films of various water-soluble alpha-, beta-, alpha-/beta-, and alpha-+beta-proteins, the amide-proton exchange, initiated by exposure of the protein film to 2H2O, has been monitored using infrared spectroscopy. The approach to obtain the kinetics of exchange for four different classes of amide protons, correlating to the different secondary structure types, has been described in detail in the preceding paper. In this work the more general applicability of the approach is illustrated by testing it for different types of proteins. The results obtained are shown not only to be comparable to reported time-resolved nuclear magnetic resonance data (as in the case of myoglobin, phospholipase A2, lysozyme, and cytochrome c), or to the more qualitative data obtained by neutron diffraction (trypsin, ribonuclease S, papain, and subtilisin BPN'), but the infrared approach us also provides with quantitative detailed insight on the distribution of exchange rate constants at the submolecular level of proteins, too complex to be studied by other techniques, as for tetrameric hemoglobin, and of proteins in which exchange is too fast to be detected by these other techniques, as is shown in this work for alpha-casein and apocytochrome c.
de Jongh
H H
HH
Laboratoire de Chimie-Physique des Macromolécules aux Interfaces, Université Libre de Bruxelles, Belgium.
Goormaghtigh
E
E
Ruysschaert
J M
JM
D000577:Amides; D000818:Animals; D001059:Apoproteins / Q000737:chemistry; D002364:Caseins / Q000737:chemistry; D002417:Cattle; D003574:Cytochrome c Group / Q000737:chemistry; D045304:Cytochromes c; D006454:Hemoglobins / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry; D010206:Papain / Q000737:chemistry; D010741:Phospholipases A / Q000737:chemistry; D054467:Phospholipases A2; D017433:Protein Structure, Secondary; D011522:Protons; D012260:Ribonucleases / Q000737:chemistry; D012995:Solubility; D017550:Spectroscopy, Fourier Transform Infrared; D013381:Subtilisins / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry; D014867:Water
D000577:Amides; D001059:Apoproteins; D002364:Caseins; D003574:Cytochrome c Group; D006454:Hemoglobins; D009211:Myoglobin; D011522:Protons; D014867:Water; D045304:Cytochromes c; D012260:Ribonucleases; D010741:Phospholipases A; D054467:Phospholipases A2; C022542:ribonuclease S; D009113:Muramidase; D013381:Subtilisins; D014357:Trypsin; D010206:Papain