1994 Fiscal Year Final Research Report Summary
Immunological Analysis of in vivo Maillard reaction of proteins
| Project/Area Number |
05806019
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
食品科学・栄養科学
|
|---|
| Research Institution | Kochi University |
|---|
Principal Investigator |
SAWAMURA Masayoshi Kochi Univ.Dept of Bioresources Sci., Professor, 農学部, 教授 (20038300)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
UKEDA Hiroyuki Kochi Uni.Dept of Bioresources Sci., Associate Professor, 農学部, 助教授 (60184991)
YAMAMOTO Sinpei Kochi Uni.Dept of Bioresources Sci., Professor, 農学部, 教授 (20036718)
|
|---|
| Project Period (FY) |
1993 – 1994
|
| Keywords | Maillard reaction / Albumin / Glutaraldehyde / Intermolecular crosslinking |
|---|
| Research Abstract |
There are some evidences showing that the bifunctional reagent glutaraldehyde (GA) might be a model compound suitable for an elucidation of post-translational modification of serum albumin. Since no report has been appeared that GA is present in serum, we assumed that a compound present in serum should give serum albumin a similar modification structure to GA.In the present project, we tried to find such a compound based on the affinity of antiserum against GA-modified rabbit serum albumin (pRSA) prepared using rabbit. The affinity with the anti-pRSA antibody was determined by ELISA.The anti-pRSA antibody showed a strong affinity only for pRSA and very low affinity for GA-modified albumins derived from other kinds of animals. The fraction of pRSA having higher molecular weight has the stronger affinity and the reduction with borohydride caused a decrease in the affinity, meaning that the antibody recognizes the polymer structure of pRSA and partly haptenic structure formed by the reaction with GA such as pyridinium. As the reaction of GA with protein can be regarded as a kind of the Maillard reaction, we examined the reaction of sugars with RSA.Among sugars examined, the modification reaction with fructose showed the highest affinity with the antibody for a given reaction time. After much longer reaction time, some affinity was observed in the modification product with glucose. The difference in the formation of the affinity clearly resulted not from the degree of modification with sugar molecules but from the extent of intermolecular crosslinking. Ascorbic acid crosslinked RSA between molecules, thereby showed the certain affinity with the antibody. The fructose-modified RSA was separated by boronate-affinity column chromatography and confirmed to inhibit competitively the interaction between pRSA and the antibody. These results suggest that intermolecular crosslinking of albumin with fructose and ascorbic acid could form a similar product to GA-modified albumin.
|
