A Simplified Method for Evaluating Chitin-Binding Activity Applied to YKL-40 (HC-gp39, CHI3L1) and Chitotriosidase
YKL-40 is structurally similar to chitotriosidase (CHIT1), an active chitinase, but it lacks chitin-degrading activity while retaining chitin-binding capability. Elevated YKL-40 levels are associated with inflammatory diseases and cancers, making it a valuable biomarker. We previously reported that...
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2024-12-01
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| author | Keita Suzuki Hidetoshi Suzuki Ami Tanaka Miwa Tanaka Kairi Takase Hiromu Takei Tomoki Kanaizumi Kazuaki Okawa Peter O. Bauer Fumitaka Oyama |
| author_facet | Keita Suzuki Hidetoshi Suzuki Ami Tanaka Miwa Tanaka Kairi Takase Hiromu Takei Tomoki Kanaizumi Kazuaki Okawa Peter O. Bauer Fumitaka Oyama |
| author_sort | Keita Suzuki |
| collection | DOAJ |
| description | YKL-40 is structurally similar to chitotriosidase (CHIT1), an active chitinase, but it lacks chitin-degrading activity while retaining chitin-binding capability. Elevated YKL-40 levels are associated with inflammatory diseases and cancers, making it a valuable biomarker. We previously reported that the W69T substitution in YKL-40 significantly reduces its chitin-binding affinity, identifying W69 as a crucial binding site. In this study, we establish a novel chitin-binding affinity evaluation method using a three-step buffer system to assess the binding strength and specificity of chitin-binding proteins and apply it to characterize YKL-40’s binding mechanism. Our findings confirm that YKL-40, through its key residue W69, exhibits highly specific and robust affinity to chitin. Unlike CHIT1, which has both a catalytic domain (CatD) and a chitin-binding domain (CBD) that allow for diverse binding and degradation activities, YKL-40 lacks a CBD and is specialized for specific chitin recognition without degrading it. Comparative analysis with YKL-39, which does not contain a corresponding W69 residue, highlights the unique role of this residue in YKL-40’s chitin-binding activity that is potentially linked to immune and inflammatory responses. Our evaluation method clarifies YKL-40’s binding properties and provides a versatile approach applicable to other chitin-binding proteins. |
| format | Article |
| id | doaj-art-37005f4ca039408e8f565a96b84198f4 |
| institution | Kabale University |
| issn | 1420-3049 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
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| series | Molecules |
| spelling | doaj-art-37005f4ca039408e8f565a96b84198f42025-01-10T13:18:37ZengMDPI AGMolecules1420-30492024-12-013011910.3390/molecules30010019A Simplified Method for Evaluating Chitin-Binding Activity Applied to YKL-40 (HC-gp39, CHI3L1) and ChitotriosidaseKeita Suzuki0Hidetoshi Suzuki1Ami Tanaka2Miwa Tanaka3Kairi Takase4Hiromu Takei5Tomoki Kanaizumi6Kazuaki Okawa7Peter O. Bauer8Fumitaka Oyama9Department of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, JapanDepartment of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, JapanDepartment of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, JapanDepartment of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, JapanDepartment of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, JapanDepartment of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, JapanDepartment of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, JapanDepartment of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, JapanBioinova a.s., Videnska 1083, 142 00 Prague, Czech RepublicDepartment of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, JapanYKL-40 is structurally similar to chitotriosidase (CHIT1), an active chitinase, but it lacks chitin-degrading activity while retaining chitin-binding capability. Elevated YKL-40 levels are associated with inflammatory diseases and cancers, making it a valuable biomarker. We previously reported that the W69T substitution in YKL-40 significantly reduces its chitin-binding affinity, identifying W69 as a crucial binding site. In this study, we establish a novel chitin-binding affinity evaluation method using a three-step buffer system to assess the binding strength and specificity of chitin-binding proteins and apply it to characterize YKL-40’s binding mechanism. Our findings confirm that YKL-40, through its key residue W69, exhibits highly specific and robust affinity to chitin. Unlike CHIT1, which has both a catalytic domain (CatD) and a chitin-binding domain (CBD) that allow for diverse binding and degradation activities, YKL-40 lacks a CBD and is specialized for specific chitin recognition without degrading it. Comparative analysis with YKL-39, which does not contain a corresponding W69 residue, highlights the unique role of this residue in YKL-40’s chitin-binding activity that is potentially linked to immune and inflammatory responses. Our evaluation method clarifies YKL-40’s binding properties and provides a versatile approach applicable to other chitin-binding proteins.https://www.mdpi.com/1420-3049/30/1/19catalytic domain (CatD)chitinchitin-binding activitychitin-binding affinity assaychitin-binding domain (CBD)chitotriosidase (CHIT1) |
| spellingShingle | Keita Suzuki Hidetoshi Suzuki Ami Tanaka Miwa Tanaka Kairi Takase Hiromu Takei Tomoki Kanaizumi Kazuaki Okawa Peter O. Bauer Fumitaka Oyama A Simplified Method for Evaluating Chitin-Binding Activity Applied to YKL-40 (HC-gp39, CHI3L1) and Chitotriosidase Molecules catalytic domain (CatD) chitin chitin-binding activity chitin-binding affinity assay chitin-binding domain (CBD) chitotriosidase (CHIT1) |
| title | A Simplified Method for Evaluating Chitin-Binding Activity Applied to YKL-40 (HC-gp39, CHI3L1) and Chitotriosidase |
| title_full | A Simplified Method for Evaluating Chitin-Binding Activity Applied to YKL-40 (HC-gp39, CHI3L1) and Chitotriosidase |
| title_fullStr | A Simplified Method for Evaluating Chitin-Binding Activity Applied to YKL-40 (HC-gp39, CHI3L1) and Chitotriosidase |
| title_full_unstemmed | A Simplified Method for Evaluating Chitin-Binding Activity Applied to YKL-40 (HC-gp39, CHI3L1) and Chitotriosidase |
| title_short | A Simplified Method for Evaluating Chitin-Binding Activity Applied to YKL-40 (HC-gp39, CHI3L1) and Chitotriosidase |
| title_sort | simplified method for evaluating chitin binding activity applied to ykl 40 hc gp39 chi3l1 and chitotriosidase |
| topic | catalytic domain (CatD) chitin chitin-binding activity chitin-binding affinity assay chitin-binding domain (CBD) chitotriosidase (CHIT1) |
| url | https://www.mdpi.com/1420-3049/30/1/19 |
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