Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K):
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Münster
2018
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| 100 | 1 | |a Kantauskaite, Marta |d 1989- |e Verfasser |0 (DE-588)1199061840 |4 aut | |
| 245 | 1 | 0 | |a Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) |c Marta Kantauskaite |
| 264 | 1 | |a Münster |c 2018 | |
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Datensatz im Suchindex
| _version_ | 1804181305757270016 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
1.
INTRODUCTION
6
1.1
RENAL
DRUG
EXCRETION
7
1.1.1
ORGANIC
CATION
TRANSPORTERS
10
1.1.2
SLC22
TRANSPORTER
FAMILY
(OCTS
AND
OCTNS)
11
1.1.3
SLC47
TRANSPORTER
FAMILY
(MATES)
12
1.2
MATE
FAMILY
CHARACTERISTICS
12
1.2.1
STRUCTURE
12
1.2.2
PHYSIOLOGICAL
SUBSTRATES
AND
INHIBITORS
14
1.2.3
SIGNIFICANCE
OF
PATHOLOGICAL
STATE
15
1.3
DETAILED
AIMS
OF
WORK
18
2.
MATERIALS
19
2.1
CELL
CULTURE
19
2.2
SOLUTIONS
AND
BUFFERS 20
2.3
CHEMICALS
21
2.4
MATERIALS
OF
MOLECULAR
BIOLOGY
METHODS
24
2.5
INSTRUMENTS
26
3.
METHODS
26
3.1
CELL
CULTURE
26
3.2
TRANSPORT
STUDIES
27
3.2.1
INFLUX
STUDIES
28
3.2.2
EFFLUX
STUDIES
28
3.2.3
DETERMINATION
OF
HALF
MAXIMAL
INHIBITORY
CONCENTRATION
29
3.3
DETERMINATION
OF
PH
USING
2
*
,7 -BIS(2-CARBOXYETHYL)-5(6)-CARBOXYFLUORESCEIN
ACETOXYMETHYL
ESTER
(BCECF-AM)
29
3.4
REGULATION
STUDIES
30
3.4.1
INFLUX
REGULATION
30
3.4.2
EFFLUX
REGULATION
31
3.5
LIVE
UPTAKE
OF
FLUORESCENT
SUBSTRATES
31
3.6
CELL
VIABILITY
TEST
31
3.7
MEASUREMENT
OF
MRNA-EXPRESSION
IN
HEK293-HMATE2K
AND
HEK293-HMATEL
CELLS
32
3.7.1
ISOLATION
OF
MRNA
32
3.7.2
REVERSE
TRANSCRIPTION
OF
CDNA
33
3.7.3
AMPLIFICATION
OF
CDNA
33
3.7.4
AGAROSE
GEL
ELECTROPHORESIS
33
3.8
STATISTICAL
ANALYSIS
34
4.
RESULTS
____________________________________________________________________________
35
4.1
VERIFICATION
OF
HMATEZK
TRANSPORTER
EXPRESSION
IN
HEK293
CELLS
USING
PCR
35
4.2
FUNCTIONAL
CHARACTERISTICS
OF
HMATEZK
35
4.2.1
INFLUX
THROUGH
HMATE2K
TRANSPORTER
35
4.2.2
EFFLUX
THROUGH
HMATE2K
TRANSPORTER
37
4.2.3
PH
DEPENDENT
TRANSPORT
ACTIVITY
39
4.3
HMATEZK
INTERACTION
WITH
NEPHROTOXIC
AGENTS
44
4.3.1
COMPETITIVE
MECHANISMS
OF
TOXIC
DRUGS
44
4.3.2
CYTOTOXICITY
STUDIES
OF
NEPHROTOXIC
AGENTS
46
4.4
SUBSTRATE
DEPENDENT
INHIBITION
OF
HMATEZK
48
4.5.
REGULATION
OF
HMATEZK
TRANSPORT
ACTIVITY
54
4.5.1
REGULATION
OF
INFLUX
54
4.5.2
REGULATION
OF
EFFLUX
59
4.6.
IMPLICATIONS
OF
TRANSPORTERS
EXPRESSED
IN
HEK293-HMATE2K
CELLS
62
4.7
LIVE
UPTAKE
OF
FLUORESCENT
SUBSTRATES
64
5.
DISCUSSION
_________________________________________________________________________
67
UNCERTAINTIES
OF
SUBSTRATE TESTING
68
UNVEILING
THE
COMPETITIVE
MECHANISM
71
THE
BURDEN
OF
RENAL
DRUG
ELIMINATION
71
RELEVANT
SINGULARITIES
OF
DRUG
CHEMICAL
STRUCTURE
72
REGULATION
OF
HMATEZK
TRANSPORTER
FUNCTION
75
TESTING
THE
EXTRUSION
THROUGH
HMATEZK
80
COORDINATED
FUNCTION
OF
HMATES
-
POSSIBLE
MECHANISM
OBSERVED
IN
IN
VITRO
STUDIES
82
6.
CONCLUSION
________________________________________________________________________
85
7.
LIST
OF
TABLES
_____________________________________________________________________
86
8.
LIST
OF
FIGURES
____________________________________________________________________
86
9.
REFERENCES
________________________________________________________________________
88
10.
CURRICULUM
VITAE
11.
ACKNOWLEDGEMENT
6
|
| adam_txt |
TABLE
OF
CONTENTS
1.
INTRODUCTION
6
1.1
RENAL
DRUG
EXCRETION
7
1.1.1
ORGANIC
CATION
TRANSPORTERS
10
1.1.2
SLC22
TRANSPORTER
FAMILY
(OCTS
AND
OCTNS)
11
1.1.3
SLC47
TRANSPORTER
FAMILY
(MATES)
12
1.2
MATE
FAMILY
CHARACTERISTICS
12
1.2.1
STRUCTURE
12
1.2.2
PHYSIOLOGICAL
SUBSTRATES
AND
INHIBITORS
14
1.2.3
SIGNIFICANCE
OF
PATHOLOGICAL
STATE
15
1.3
DETAILED
AIMS
OF
WORK
18
2.
MATERIALS
19
2.1
CELL
CULTURE
19
2.2
SOLUTIONS
AND
BUFFERS 20
2.3
CHEMICALS
21
2.4
MATERIALS
OF
MOLECULAR
BIOLOGY
METHODS
24
2.5
INSTRUMENTS
26
3.
METHODS
26
3.1
CELL
CULTURE
26
3.2
TRANSPORT
STUDIES
27
3.2.1
INFLUX
STUDIES
28
3.2.2
EFFLUX
STUDIES
28
3.2.3
DETERMINATION
OF
HALF
MAXIMAL
INHIBITORY
CONCENTRATION
29
3.3
DETERMINATION
OF
PH
USING
2
*
,7'-BIS(2-CARBOXYETHYL)-5(6)-CARBOXYFLUORESCEIN
ACETOXYMETHYL
ESTER
(BCECF-AM)
29
3.4
REGULATION
STUDIES
30
3.4.1
INFLUX
REGULATION
30
3.4.2
EFFLUX
REGULATION
31
3.5
LIVE
UPTAKE
OF
FLUORESCENT
SUBSTRATES
31
3.6
CELL
VIABILITY
TEST
31
3.7
MEASUREMENT
OF
MRNA-EXPRESSION
IN
HEK293-HMATE2K
AND
HEK293-HMATEL
CELLS
32
3.7.1
ISOLATION
OF
MRNA
32
3.7.2
REVERSE
TRANSCRIPTION
OF
CDNA
33
3.7.3
AMPLIFICATION
OF
CDNA
33
3.7.4
AGAROSE
GEL
ELECTROPHORESIS
33
3.8
STATISTICAL
ANALYSIS
34
4.
RESULTS
_
35
4.1
VERIFICATION
OF
HMATEZK
TRANSPORTER
EXPRESSION
IN
HEK293
CELLS
USING
PCR
35
4.2
FUNCTIONAL
CHARACTERISTICS
OF
HMATEZK
35
4.2.1
INFLUX
THROUGH
HMATE2K
TRANSPORTER
35
4.2.2
EFFLUX
THROUGH
HMATE2K
TRANSPORTER
37
4.2.3
PH
DEPENDENT
TRANSPORT
ACTIVITY
39
4.3
HMATEZK
INTERACTION
WITH
NEPHROTOXIC
AGENTS
44
4.3.1
COMPETITIVE
MECHANISMS
OF
TOXIC
DRUGS
44
4.3.2
CYTOTOXICITY
STUDIES
OF
NEPHROTOXIC
AGENTS
46
4.4
SUBSTRATE
DEPENDENT
INHIBITION
OF
HMATEZK
48
4.5.
REGULATION
OF
HMATEZK
TRANSPORT
ACTIVITY
54
4.5.1
REGULATION
OF
INFLUX
54
4.5.2
REGULATION
OF
EFFLUX
59
4.6.
IMPLICATIONS
OF
TRANSPORTERS
EXPRESSED
IN
HEK293-HMATE2K
CELLS
62
4.7
LIVE
UPTAKE
OF
FLUORESCENT
SUBSTRATES
64
5.
DISCUSSION
_
67
UNCERTAINTIES
OF
SUBSTRATE TESTING
68
UNVEILING
THE
COMPETITIVE
MECHANISM
71
THE
BURDEN
OF
RENAL
DRUG
ELIMINATION
71
RELEVANT
SINGULARITIES
OF
DRUG
CHEMICAL
STRUCTURE
72
REGULATION
OF
HMATEZK
TRANSPORTER
FUNCTION
75
TESTING
THE
EXTRUSION
THROUGH
HMATEZK
80
COORDINATED
FUNCTION
OF
HMATES
-
POSSIBLE
MECHANISM
OBSERVED
IN
IN
VITRO
STUDIES
82
6.
CONCLUSION
_
85
7.
LIST
OF
TABLES
_
86
8.
LIST
OF
FIGURES
_
86
9.
REFERENCES
_
88
10.
CURRICULUM
VITAE
11.
ACKNOWLEDGEMENT
6 |
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| author | Kantauskaite, Marta 1989- |
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| institution | BVB |
| language | English |
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| spelling | Kantauskaite, Marta 1989- Verfasser (DE-588)1199061840 aut Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) Marta Kantauskaite Münster 2018 100 Blätter Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Westfälische Wilhelms-Universität Münster 2019 Beschränkt für den Austausch (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1198900628/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032036058&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Kantauskaite, Marta 1989- Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) |
| subject_GND | (DE-588)4113937-9 |
| title | Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) |
| title_auth | Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) |
| title_exact_search | Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) |
| title_exact_search_txtP | Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) |
| title_full | Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) Marta Kantauskaite |
| title_fullStr | Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) Marta Kantauskaite |
| title_full_unstemmed | Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) Marta Kantauskaite |
| title_short | Functional characterization of human multidrug and toxin extrusion transporter 2K (hMATE2K) |
| title_sort | functional characterization of human multidrug and toxin extrusion transporter 2k hmate2k |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1198900628/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032036058&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT kantauskaitemarta functionalcharacterizationofhumanmultidrugandtoxinextrusiontransporter2khmate2k |
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