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267449 occurrences (No.100 in the rank) during 5 years in the PubMed. [no cache] 500 found

211) In the present study, we have used alanine-scanning mutagenesis to explore the structural and functional relationships within this loop of the yeast plasma membrane Pma1 H(+)-ATPase.

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PMID:24256122 DOI:10.1080/07391102.2013.849619

2015 Journal of biomolecular structure & dynamics

* Point mutations in the extracytosolic loop between transmembrane segments M5 and M6 of the yeast Pma1 H+-ATPase: alanine-scanning mutagenesis.

- Membrane-spanning segments M4, M5, M6, and M8 of the H(+)-, Ca(2+)-, and K(+), Na(+)-ATPases, which belong to the P2-type pumps are the core through which cations are transported. M5 and M6 loop is a short extracytoplasmic stretch of the seven amino acid residues (714-DNSLDID) connecting two of these segments, M5 and M6, where residues involved in the formation of the proton-binding site(s) are located. In the present study, we have used alanine-scanning mutagenesis to explore the structural and functional relationships within this loop of the yeast plasma membrane Pma1 H(+)-ATPase. Of the 7 Ala mutants made, substitution for the most conserved residue (Leu-717) has led to a severe misfolding and complete block in biogenesis of the mutant enzyme. The replacement of Asp-714 has also caused misfolding leading to significant decrease in the expression of the mutant and loss of activity. The remaining mutants were expressed in secretory vesicles at 21-119% of the wild-type level and were active enough to be analyzed in detail. One of these mutants (I719A) showed five- to threefold decrease in both expression and ATP hydrolyzing and H(+) pumping activities and also threefold reduction in the coupling ratio between ATP hydrolysis and H(+) transport. Thus, Ala substitutions at three positions of the seven seriously affected biogenesis, folding, stability and/or functioning of the enzyme. Taken together, these results lead to suggestion that M5 and M6 loop play an important role in the protein stability and function and is responsible for proper arrangement of transmembrane segments M5 and M6 and probably other domains of the enzyme. Results for additional conserved substitutions (Asn and Glu) at Asp-714 and Asp-720 confirmed this suggestion.

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(1)50 magnetic	(18)4 gastrointestinal	(35)3 integration	(52)2 connections
(2)38 connectivity	(19)4 outcome	(36)3 interactions	(53)2 differentiation
(3)29 and	(20)4 outcomes	(37)3 movement	(54)2 domains
(4)8 limitations	(21)4 relationships	(38)3 single	(55)2 endurance
(5)7 network	(22)4 unit	(39)3 status	(56)2 health
(6)7 recovery	(23)3 Assessment	(40)3 status,	(57)2 imaging
(7)7 role	(24)3 MRI	(41)3 studies	(58)2 impact
(8)6 changes	(25)3 Well-Being	(42)2 CD133	(59)2 integrity
(9)6 results	(26)3 activation	(43)2 HL	(60)2 losses
(10)5 alterations	(27)3 brain	(44)2 ability	(61)2 networks
(11)5 limitation	(28)3 consequences	(45)2 activity	(62)2 neuronal
(12)5 neuroimaging	(29)3 coupling	(46)2 analyses	(63)2 organization
(13)5 properties	(30)3 decline	(47)2 as	(64)2 parameters
(14)5 theory	(31)3 deficits	(48)2 assays	(65)2 roles
(15)4 capacity	(32)3 differences	(49)2 assessment	(66)2 significance
(16)4 connectivity,	(33)3 impairment	(50)2 characteristics	(67)2 specialization
(17)4 foods	(34)3 improvement	(51)2 cognitive

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--- WordNet output for functional --- =>機能の, 関数の, 機能する, 機能上の, 実用的な, 機能的な Overview of adj functional The adj functional has 6 senses (first 2 from tagged texts) 1. (3) functional -- (designed for or capable of a particular function or use; "a style of writing in which every word is functional"; "functional architecture") 2. (1) functional -- (involving or affecting function rather than physiology; "functional deafness") 3. functional -- (relating to or based on function especially as opposed to structure; "the problem now is not a constitutional one; it is a functional one"; "delegates elected on a functional rather than a geographical basis") 4. functional, usable, useable, operable, operational -- (fit or ready for use or service; "the toaster was still functional even after being dropped"; "the lawnmower is a bit rusty but still usable"; "an operational aircraft"; "the dishwasher is now in working order") 5. functional -- (designed for or adapted to a function or use; "functional education selects knowledge that is concrete and usable rather than abstract and theoretical"; "functional architecture") 6. running, operative, functional, working -- ((of e.g. a machine) performing or capable of performing; "in running (or working) order"; "a functional set of brakes") --- WordNet end ---