CN109678928A - A kind of tripeptides with 1 inhibitory activity of cholinesterase and beta amyloid precursor protein cleavage enzyme - Google Patents
A kind of tripeptides with 1 inhibitory activity of cholinesterase and beta amyloid precursor protein cleavage enzyme Download PDFInfo
- Publication number
- CN109678928A CN109678928A CN201811655328.1A CN201811655328A CN109678928A CN 109678928 A CN109678928 A CN 109678928A CN 201811655328 A CN201811655328 A CN 201811655328A CN 109678928 A CN109678928 A CN 109678928A
- Authority
- CN
- China
- Prior art keywords
- wir
- tripeptides
- cholinesterase
- precursor protein
- cleavage enzyme
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0821—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Life Sciences & Earth Sciences (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention belongs to technical field of polypeptide, and in particular to a kind of tripeptides with 1 inhibitory activity of cholinesterase and beta amyloid precursor protein cleavage enzyme, amino acid sequence are Trp-Ile-Arg (WIR).Active kyrine WIR derives from food proteins, with high security, it is cheap, can industrialization outstanding advantages.Active kyrine WIR or derivatives thereof can be used for preparing senile dementia treatment/prophylactic agent, or as function food additive, for old dementia patients long-term treatment health care, be with a wide range of applications and highly important meaning.
Description
Technical field
The invention belongs to technical field of polypeptide, and in particular to one kind has cholinesterase and beta amyloid precursor protein cleavage
The tripeptides of 1 inhibitory activity of enzyme.
Background technique
Since the elderly population number in the whole world increases, dementia becomes the maximum global public health that current people face
One of challenge.Alzheimer disease (Alzheimer ' s disease, AD) is the most common dementia type, main feature are as follows: big
Brain gradually degenerate cause the loss of memory, get lost, anxiety, vain hope, depression, insomnia, roaming, learning disorder and cognitive function decline
It moves back.The study found that loss, β-shallow lake caused by many factors lead to the development of this disease of AD, including levels of acetylcholine reduces
The increase etc. of powder sample albumen.Although caused by AD is multifactor, pathogenic factor understands there is presently no complete, currently used
The most common treatment method is by acetylcholine esterase inhibition (AChE) Lai Shixian.AChE is one kind at cholinergic synapse
Hydrolysis neurotransmitter acetylcholine is to terminate the enzyme of neurotransmission.Some studies pointed out that another unspecific cholinesterases ---
Butyrylcholine esterase (BChE), and the target spot for the treatment of AD；Simultaneously also some researches show that the generation of AD and beta-amyloid protein with
Neurotoxicity is damaged caused by accumulation related.Clinical test shows β-shallow lake of most patients with Alzheimer disease
1 (BACE1) expression of powder sample precursor protein cleavage enzyme, which has, to be increased, so as to cause more beta-amyloid proteins are generated.Cause
This, acquisition, which has both, inhibits cholinesterase (ChE) and the active active constituent of BACE1 that will become prevention and treatment alzheimer's disease
Effective ways.
Biologically active peptide be in protein 20 natural amino acids with it is different form and arrangement mode constituted slave dipeptides to
Complicated linear, ring structure different peptides, especially some oligopeptides not only have digestion and absorption more better than protein
Performance, while also having the effects that adjust human physiological functions.Thus biologically active peptide has important in functional food
Application value.Therefore, the peptides of cholinesterase (ChE) and beta amyloid precursor protein cleavage enzyme 1 (BACE1) inhibitory activity are had both
The following effective component for preventing and treating Alzheimer disease will be become.
Summary of the invention
It is an object of the present invention to provide the activity to AChE, BChE, BACE1 to have continual and steady inhibiting effect, has good
The tripeptides of good anti-AD effect, can be applied to food, health care product and field of biological pharmacy.
Tripeptides with 1 inhibitory activity of cholinesterase and beta-amyloid protein precursor protein cleavage enzyme of the invention, ammonia
Base acid sequence is WIR.
The tripeptides of 1 inhibitory activity of cholinesterase and beta-amyloid protein precursor protein cleavage enzyme of aforementioned present invention should wrap
It includes different by amino acid fine jade luminosity and is defined as the tripeptides of the various configuration of L-type amino acid or D type amino acid.
Preferably, the three of 1 inhibitory activity of cholinesterase and beta-amyloid protein precursor protein cleavage enzyme of the present invention
Peptide is originated from fish-egg albumen.
The tripeptides of 1 inhibitory activity of cholinesterase and beta-amyloid protein precursor protein cleavage enzyme of the invention has good
AChE, BACE1 inhibitory effect, IC50 value are respectively 0.02mg/mL and 0.96mg/mL, when WIR concentration is 0.5mg/mL pair
The active inhibiting rate of BChE is 33.5%.
The purpose of the present invention is achieved through the following technical solutions:
(1) the virtual enzymatic hydrolysis and virtual screening of protein sequence
The present invention is by ExPASy PeptideCutter (http://web.expasy.org/peptide_cutter/)
Online virtual digestion tool virtually digests fish-egg protein sequence, the tripeptide sequence and BIOPEP that enzymatic hydrolysis is generated
(http://www.uwm.edu.pl/biochemia/index.php/en/biopep) is compared, screening acquire without
The tripeptides of report.By online tool ToxinPred, peptide property calculator, PeptideRanker and
AdmetSAR carries out toxicity, water solubility, bioactivity and ADME to the tripeptide sequence without report and (absorbs, distribution, metabolism, row
Let out) prediction of property.Screening obtains three of Activity Score higher than 0.5, nontoxic, good water solubility and with good small intestine permeability
Peptide.
(2) targeting screening
From acquisition AChE (1EVE), BChE (1POI), BACE1 in PDB database (http://www.rcsb.org/)
(1TQF), and as protein target, molecule pair is carried out by the CDOCKER program of Discovery Studio (DS) 2017
Fetch the tripeptides that screening can combine closely with AChE, BChE, BACE1.It is obtained according to ' CDOCKER-INIERACTION-ENERGY '
Divide, form the number of hydrogen bond and the key amino acid of effect, screening has the three of potential AChE, BChE, BACE1 inhibitory activity
Peptide.
(3) external AChE, BChE and BACE1 inhibitory activity measurement
It is verified by the external inhibitory activity of ChE of the spectrophotometry to tripeptides.Use acetylcholine (ACh), butyryl
Choline (BCh) is respectively as substrate, by sodium phosphate buffer (pH 8.0), test sample solution and the mixing of AChE/BChE solution
And 15min is being incubated at 37 DEG C.Then DTNB and ACh/BCh (10mmol/L) is added to mixture to be sufficiently mixed uniformly, 37
DEG C heat preservation 30min after.Each group absorbance value is detected with microplate reader at 412nm.
Use the BACE1 inhibitory activity of fluorescence spectrophotometry tripeptides.Take 75-X μ L fluoremetry buffer (PH
4.5) mixture of, 20 μ LBACE1 substrate solution (50 μM), 5 μ L BACE1 enzymes (0.3u/ μ L) and X μ L test sample is 37
It is incubated for 2 hours at DEG C, is added with 80 μ L buffers and 20 μ L BACE1 substrate solutions and 75 μ L buffers and BACE1 substrate solution
5 μ L BACE1 enzymes (0.3u/ μ L) are used as blank control, pass through its activity of fluorescence detection.
The present invention having the beneficial effect that compared with prior art
Screening has obtained that AChE, BChE and BACE1 active three can be inhibited simultaneously the present invention from fish-egg albumen for the first time
Peptide WIR, specifies the structure of tripeptides WIR, and illustrates tripeptides WIR and AChE, the mechanism of action of BChE and BACE1.Therefore three
Peptide WIR is used in the food with adjuvant treatment senile dementia, health care product and drug as functional component, is had good latent
Power and application prospect.
Detailed description of the invention
3 width of attached drawing of the present invention, in which:
Figure 1A ChE-WIR (PDB ID:1EVE) interaction；
Fig. 2 BChE-WIR (PDB ID:1POI) interaction；
Fig. 3 BACE1-WIR (PDB ID:1TQF) interaction.
Specific embodiment
The invention will be further elaborated in a manner of specific embodiment below.
The virtual enzymatic hydrolysis and virtual screening of 1 atlantic salmon fish-egg albumin A ChE peptide for inhibiting of embodiment
Based in sequence of threads ExPASy PeptideCutter, two kinds of typical pipe intestinal digesting protease, that is, stomach eggs are used
White enzyme (EC 3.4.23.1) and trypsase (EC 3.4.21.4) carry out simulation enzymatic hydrolysis, ammonia to atlantic salmon fish-egg albumen
Base acid sequence is mrttaafllvlcllaishawdcqpvvdiknlmqidaglgqvvatdt sqipyylvgdkwirmpgyl
khitvgpagiwgvnkdyaiykyvagnwvqaagllkqldaggdqfvvganmndspfclassatvgykgpgsplpwtg
lpgsvkyyscgpfgcwavnnnddiylmsl nqdcqnngwshiegklsmievatdgsvfgvnsvgsvytrdgitask
pegtgwsnipmcmq mkhvtydlgrlwvisksgftmvcth.Pass through online tool ToxinPred, Peptide
Property calculator, PeptideRanker and admetSAR carry out toxicity, water-soluble to the tripeptide sequence without report
The prediction of property, bioactivity and ADME property.Screening obtain nontoxic, dissolubility is good, Activity Score be higher than 0.5 and have it is good
The tripeptides of small intestine permeability.
The targeting screening of embodiment 2
Respectively with AChE (PDB ID:1EVE), BChE (PDB ID:1POI) and BACE1 (PDB ID:1TQF) are protein targets
Mark carries out molecular docking by the CDOCKER program of Discovery Studio (DS) 2017 and screens to obtain energy and AChE, BChE
The tripeptides that BACE1 combines closely, the key amino acid for scoring and acting in conjunction with CDOCKER, screening have obtained theoretically having latent
In the active tripeptides WIR of anti-AD.The result shows that WIR can with key residues, that is, Ser286, Tyr334 of AChE, Phe288,
Tyr121, Trp279 and Tyr70 combine (Fig. 1), tie with key residues, that is, His438, Trp82, Tyr332 and Asp70 of BChE
Close (Fig. 2), with key residues, that is, Ile110, Gly34 of BACE1, Leu30, Trp115, Thr231, Thr72, Gly34,
Arg235, Gly230, Asp32, Asp228, Asn233 and Lys321 combine (Fig. 3).Can add it is some it is specific scoring or
The analysis of the hydrogen bond of formation.Therefore, tripeptides WIR is synthesized, ESI-MS measure molecular weight be 473.58Da ([M+H]+
474.00Da。
The 3 external AChE of tripeptides WIR of embodiment, the identification of BChE and BACE1 inhibitory activity
Itself AChE and BChE activity is verified by fluorescence spectrophotometry.The result shows that WIR can be effectively suppressed
The activity of AChE, IC50Value is respectively 0.02mg/mL, and when WIR concentration is 0.5mg/mL, the inhibiting rate to BChE is
33.5%.BACE1 inhibitory activity is verified by fluorescence spectrophotometry, the results showed that tripeptides WIR can be effectively suppressed
The activity of BACE1, IC50Value is 0.96mg/mL.
Technical solution of the present invention is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to restrict the invention, it should be pointed out that: for those skilled in the art
For, it under the premise without departing from the spirit and scope of the present invention, can also make a variety of changes and modification, these variations and become
Type also should be regarded as protection scope of the present invention.
Claims (3)
1. a kind of tripeptides with 1 inhibitory activity of cholinesterase and beta amyloid precursor protein cleavage enzyme, which is characterized in that amino
Acid sequence is Trp-Ile-Arg (WIR).
2. according to claim 1 have inhibition cholinesterase and the active tripeptides of beta amyloid precursor protein cleavage enzyme 1,
It is characterized in that WIR inhibits the active IC of AChE, BACE150Value is respectively 0.02mg/mL and 0.96mg/mL, when WIR concentration is
It is 33.5% to the active inhibiting rate of BChE when 0.5mg/mL.
3. according to claim 1 have inhibition cholinesterase and beta-amyloid protein precursor protein cleavage enzyme 1 active
Tripeptides, which is characterized in that the amino acid residue of catalytic site that WIR and AChE interaction is related to be respectively Ser286,
Tyr334, Phe288, Tyr121, Trp279 and Tyr70；The amino acid for the catalytic site that WIR and BChE interaction is related to is residual
Base is respectively His438, Trp82, Tyr332 and Asp70；The amino acid for the catalytic site that WIR and BACE1 interaction is related to is residual
Base be respectively Ile110, Gly34, Leu30, Trp115, Thr231, Thr72, Gly34, Arg235, Gly230, Asp32,
Asp228, Asn233 and Lys321.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811655328.1A CN109678928A (en) | 2018-12-27 | 2018-12-27 | A kind of tripeptides with 1 inhibitory activity of cholinesterase and beta amyloid precursor protein cleavage enzyme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811655328.1A CN109678928A (en) | 2018-12-27 | 2018-12-27 | A kind of tripeptides with 1 inhibitory activity of cholinesterase and beta amyloid precursor protein cleavage enzyme |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109678928A true CN109678928A (en) | 2019-04-26 |
Family
ID=66191740
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811655328.1A Pending CN109678928A (en) | 2018-12-27 | 2018-12-27 | A kind of tripeptides with 1 inhibitory activity of cholinesterase and beta amyloid precursor protein cleavage enzyme |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109678928A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110317240A (en) * | 2019-07-02 | 2019-10-11 | 渤海大学 | A kind of tripeptides DMG and its application |
| CN112940075A (en) * | 2021-01-07 | 2021-06-11 | 渤海大学 | Acetylcholine esterase inhibitory peptide and application thereof |
| CN117384239A (en) * | 2023-12-12 | 2024-01-12 | 深圳保时健生物工程有限公司 | Anti-aging salmon roe tripeptide as well as preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104293794A (en) * | 2014-09-24 | 2015-01-21 | 南方医科大学 | Nucleic acid aptamers specifically combined with beta-amyloid precursor protein lyase 1 and application of aptamers |
| CN107132360A (en) * | 2017-05-08 | 2017-09-05 | 南京中医药大学 | Active peptides high-throughput screening method based on tandem mass spectrum and molecular docking |
| CN108042790A (en) * | 2017-12-20 | 2018-05-18 | 渤海大学 | A kind of pentapeptide KLPGF inhibits the application of AChE activity |
-
2018
- 2018-12-27 CN CN201811655328.1A patent/CN109678928A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104293794A (en) * | 2014-09-24 | 2015-01-21 | 南方医科大学 | Nucleic acid aptamers specifically combined with beta-amyloid precursor protein lyase 1 and application of aptamers |
| CN107132360A (en) * | 2017-05-08 | 2017-09-05 | 南京中医药大学 | Active peptides high-throughput screening method based on tandem mass spectrum and molecular docking |
| CN108042790A (en) * | 2017-12-20 | 2018-05-18 | 渤海大学 | A kind of pentapeptide KLPGF inhibits the application of AChE activity |
Non-Patent Citations (4)
| Title |
|---|
| CHEONGYUN JANG等: "Identification of novel acetylcholinesterase inhibitors designed by pharmacophorebased virtual screening, molecular docking and bioassay", 《SCIENTIFIC REPORTS》 * |
| CHEONGYUN JANG等: "Identification of novel acetylcholinesterase inhibitors designed by pharmacophorebased virtual screening, molecular docking and bioassay", 《SCIENTIFIC REPORTS》, vol. 8, 8 October 2018 (2018-10-08), pages 14921 * |
| SHENGQI RAO等: "ACE inhibitory peptides and antioxidant peptides derived from in vitro digestion hydrolysate of hen egg white lysozyme", 《FOOD CHEMISTRY》 * |
| SHENGQI RAO等: "ACE inhibitory peptides and antioxidant peptides derived from in vitro digestion hydrolysate of hen egg white lysozyme", 《FOOD CHEMISTRY》, vol. 135, 19 May 2012 (2012-05-19), pages 1250 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110317240A (en) * | 2019-07-02 | 2019-10-11 | 渤海大学 | A kind of tripeptides DMG and its application |
| CN112940075A (en) * | 2021-01-07 | 2021-06-11 | 渤海大学 | Acetylcholine esterase inhibitory peptide and application thereof |
| CN112940075B (en) * | 2021-01-07 | 2023-03-31 | 渤海大学 | Acetylcholine esterase inhibitory peptide and application thereof |
| CN117384239A (en) * | 2023-12-12 | 2024-01-12 | 深圳保时健生物工程有限公司 | Anti-aging salmon roe tripeptide as well as preparation method and application thereof |
| CN117384239B (en) * | 2023-12-12 | 2024-02-09 | 深圳保时健生物工程有限公司 | Anti-aging salmon roe tripeptide as well as preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yu et al. | Identification and molecular docking study of a novel angiotensin-I converting enzyme inhibitory peptide derived from enzymatic hydrolysates of Cyclina sinensis | |
| Majumder et al. | Molecular targets of antihypertensive peptides: understanding the mechanisms of action based on the pathophysiology of hypertension | |
| Kim et al. | Beneficial effects of soybean-derived bioactive peptides | |
| CN109678928A (en) | A kind of tripeptides with 1 inhibitory activity of cholinesterase and beta amyloid precursor protein cleavage enzyme | |
| Zheng et al. | Isolation of novel ACE-inhibitory and antioxidant peptides from quinoa bran albumin assisted with an in silico approach: Characterization, in vivo antihypertension, and molecular docking | |
| Darewicz et al. | Angiotensin I-converting enzyme (ACE) inhibitory activity and ACE inhibitory peptides of salmon (Salmo salar) protein hydrolysates obtained by human and porcine gastrointestinal enzymes | |
| Bhatia et al. | Introduction to enzymes and their applications | |
| Liu et al. | Purification, characterization and evaluation of inhibitory mechanism of ACE inhibitory peptides from pearl oyster (Pinctada fucata martensii) meat protein hydrolysate | |
| Manzanares et al. | Improving health-promoting effects of food-derived bioactive peptides through rational design and oral delivery strategies | |
| Zheng et al. | Preparation, identification, molecular docking study and protective function on HUVECs of novel ACE inhibitory peptides from protein hydrolysate of skipjack tuna muscle | |
| CN104540960B (en) | The manufacture method of collagen peptide composition, 4 Inhibitorses of DPP and blood glucose value raise inhibitor | |
| CN104349768A (en) | Compositions comprising apomorphine and organic acids and uses thereof | |
| Kim et al. | Current perspectives on the physiological activities of fermented soybean-derived cheonggukjang | |
| Feng et al. | Walnut protein hydrolysates play a protective role on neurotoxicity induced by d-galactose and aluminum chloride in mice | |
| Zhang et al. | Purification, identification, activity evaluation, and stability of antioxidant peptides from alcalase hydrolysate of Antarctic Krill (Euphausia superba) proteins | |
| Lin et al. | Antioxidative peptides derived from enzyme hydrolysis of bone collagen after microwave assisted acid pre-treatment and nitrogen protection | |
| Piotrowicz et al. | Antioxidant, angiotensin-converting enzyme inhibitory properties and blood-pressure-lowering effect of rice bran protein hydrolysates | |
| Tałałaj et al. | The effects of a novel series of KTTKS analogues on cytotoxicity and proteolytic activity | |
| Yang et al. | Accumulation and identification of angiotensin-converting enzyme inhibitory peptides from wheat germ | |
| Zhang et al. | Separation and lipid inhibition effects of a novel Decapeptide from Chlorella pyenoidose | |
| Fan et al. | A study on the skin whitening activity of digesta from edible bird’s nest: a mucin glycoprotein | |
| CN103146789B (en) | Preparation method of collagen | |
| Ngoc et al. | Insights into Bioactive Peptides in Cosmetics | |
| Fadimu et al. | In-silico analysis and antidiabetic effect of α-amylase and α-glucosidase inhibitory peptides from lupin protein hydrolysate: Enzyme-peptide interaction study using molecular docking approach | |
| Lu et al. | Alpha-Glucosidase Inhibitory Peptides: Sources, Preparations, Identifications, and Action Mechanisms |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |