P44308 (NADR_HAEIN) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Bifunctional NAD biosynthesis protein NadR

Including the following 2 domains:

1. Nicotinamide mononucleotide adenylyltransferase
   Short name=NMN adenylyltransferase
   Short name=NMN-AT
   Short name=NMNAT
   EC=2.7.7.1
   Alternative name(s):
   Nicotinamide ribonucleotide adenylyltransferase
   Nicotinamide-nucleotide adenylyltransferase
2. Ribosylnicotinamide kinase
   Short name=RNK
   EC=2.7.1.22
   Alternative name(s):
   Nicotinamide riboside kinase
   Short name=NRK
   Short name=NmR-K

Gene names

Name:	nadR
Ordered Locus Names:	HI_0763

Organism

Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)

Taxonomic identifier

71421 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Pasteurellales › Pasteurellaceae › Haemophilus

Protein attributes

Sequence length	421 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	This enzyme has two activities: nicotinamide mononucleotide (NMN) adenylyltransferase and ribosylnicotinamide (RN) kinase. The RN kinase activity catalyzes the phosphorylation of RN to form nicotinamide ribonucleotide. The NMN adenylyltransferase activity catalyzes the transfer of the AMP moiety of ATP to nicotinamide ribonucleotide to form NAD+. Ref.2
Catalytic activity	ATP + nicotinamide ribonucleotide = diphosphate + NAD+. Ref.2 ATP + N-ribosylnicotinamide = ADP + nicotinamide ribonucleotide. Ref.2
Enzyme regulation	Feed-back regulated by NAD. At high levels of NAD the RN kinase activity is inhibited.
Pathway	Cofactor biosynthesis; NAD(+) biosynthesis. [regulation] Cofactor biosynthesis; NAD(+) biosynthesis; NAD(+) from nicotinamide D-ribonucleotide: step 1/1.
Subunit structure	Homotetramer. Ref.4
Subcellular location	Cell membrane; Peripheral membrane protein. Cytoplasm. Note: Found as a soluble cytoplasmic protein as well as a membrane-associated protein. In combination with corepressor (NAD), the cytoplasmic form of NadR would be capable of acting as a transcriptional repressor. Ref.3
Sequence similarities	In the N-terminal section; belongs to the bacterial NMN adenylyltransferase family. In the C-terminal section; belongs to the bacterial RNK family.
Biophysicochemical properties	Kinetic parameters: KM=0.14 mM for NMN Ref.2

Ontologies

Keywords
Biological process	Pyridine nucleotide biosynthesis
Cellular component	Cell membrane Cytoplasm Membrane
Coding sequence diversity	Alternative initiation
Ligand	ATP-binding NAD Nucleotide-binding
Molecular function	Kinase Transferase
Technical term	3D-structure Complete proteome Multifunctional enzyme Reference proteome
Gene Ontology (GO)
Biological process	negative regulation of transcription, DNA-dependent Inferred from electronic annotation. Source: InterPro pyridine nucleotide biosynthetic process Inferred from electronic annotation. Source: UniProtKB-KW transport Inferred from electronic annotation. Source: InterPro
Cellular component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell plasma membrane Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	ATP binding Inferred from electronic annotation. Source: UniProtKB-KW DNA binding Inferred from electronic annotation. Source: InterPro nicotinamide-nucleotide adenylyltransferase activity Inferred from electronic annotation. Source: EC ribosylnicotinamide kinase activity Inferred from electronic annotation. Source: EC
Complete GO annotation...

Alternative products

This entry describes 2 isoforms produced by alternative initiation. [Align] [Select]

Isoform Long (identifier: P44308-1)

This isoform has been chosen as the 'canonical' sequence. All positional information in this entry refers to it. This is also the sequence that appears in the downloadable versions of the entry.

Isoform Short (identifier: P44308-2)

The sequence of this isoform differs from the canonical sequence as follows:
     1-51: Missing.

Note: Shows the same catalytic activity as isoform Long.

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 421

421

Bifunctional NAD biosynthesis protein NadR

PRO_0000096689

Regions

Nucleotide binding

64 – 67

4

NAD 1

Nucleotide binding

139 – 152

14

NAD 1

Nucleotide binding

172 – 174

3

NAD 1

Nucleotide binding

199 – 201

3

NAD 1

Nucleotide binding

254 – 256

3

NAD 2

Nucleotide binding

289 – 292

4

NAD 2

Region

57 – 224

168

Nicotinamide mononucleotide adenylyltransferase

Region

225 – 421

197

Ribosylnicotinamide kinase

Sites

Binding site

71

1

NAD 1

Binding site

98

1

NAD 1

Natural variations

Alternative sequence

1 – 51

51

Missing in isoform Short.

VSP_040071

Experimental info

Mutagenesis

126

1

K → A or T: Significant reduction of RNK activity, also NMN adenylyltransferase activity is impaired. Ref.3

Mutagenesis

238

1

G → N or S: Complete loss of RNK activity, no effect on NMN adenylyltransferase activity. Ref.3

Mutagenesis

256

1

W → F: No effect on enzyme activities, but NAD feedback inhibition is almost lost. Ref.3

Mutagenesis

292

1

Y → I: Almost no effect. Ref.3

Mutagenesis

304

1

D → C, N or S: Complete loss of RNK activity, no effect on NMN adenylyltransferase activity. Ref.3

Mutagenesis

352

1

R → A, C, M or N: Complete loss of RNK activity, no effect on NMN adenylyltransferase activity. Ref.3

Secondary structure

1

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421

Helix Strand Turn

Details...

Beta strand

60 – 65

6

Helix

72 – 82

11

Beta strand

86 – 94

9

Helix

96 – 105

10

Helix

114 – 124

11

Turn

127 – 131

5

Beta strand

132 – 138

7

Beta strand

140 – 142

3

Helix

149 – 162

14

Beta strand

168 – 171

4

Helix

175 – 177

3

Helix

178 – 184

7

Beta strand

188 – 190

3

Helix

203 – 208

6

Helix

210 – 216

7

Turn

219 – 221

3

Helix

222 – 224

3

Beta strand

227 – 232

6

Helix

238 – 249

12

Beta strand

253 – 255

3

Helix

260 – 265

6

Beta strand

266 – 269

4

Turn

276 – 278

3

Helix

279 – 296

18

Beta strand

298 – 305

8

Helix

307 – 318

12

Helix

323 – 331

9

Beta strand

335 – 341

7

Helix

360 – 372

13

Helix

373 – 375

3

Beta strand

379 – 382

4

Helix

386 – 400

15

Sequences

	Sequence		Length	Mass (Da)
	Isoform Long [UniParc]. Last modified November 1, 1995. Version 1. Checksum: D9A4FD4970A6E7E8	FASTA	421	49,432
	10 20 30 40 50 60 MGFTTGREFH PALRMRAKYN AKYLGTKSER EKYFHLAYNK HTQFLRYQEQ IMSKTKEKKV 70 80 90 100 110 120 GVIFGKFYPV HTGHINMIYE AFSKVDELHV IVCSDTVRDL KLFYDSKMKR MPTVQDRLRW 130 140 150 160 170 180 MQQIFKYQKN QIFIHHLVED GIPSYPNGWQ SWSEAVKTLF HEKHFEPSIV FSSEPQDKAP 190 200 210 220 230 240 YEKYLGLEVS LVDPDRTFFN VSATKIRTTP FQYWKFIPKE ARPFFAKTVA ILGGESSGKS 250 260 270 280 290 300 VLVNKLAAVF NTTSAWEYGR EFVFEKLGGD EQAMQYSDYP QMALGHQRYI DYAVRHSHKI 310 320 330 340 350 360 AFIDTDFITT QAFCIQYEGK AHPFLDSMIK EYPFDVTILL KNNTEWVDDG LRSLGSQKQR 370 380 390 400 410 420 QQFQQLLKKL LDKYKVPYIE IESPSYLDRY NQVKAVIEKV LNEEEISELQ NTTFPIKGTS Q
	Isoform Short [UniParc]. Checksum: 6F99B0CA15ABBB59	FASTA	370	43,192
	10 20 30 40 50 60 MSKTKEKKVG VIFGKFYPVH TGHINMIYEA FSKVDELHVI VCSDTVRDLK LFYDSKMKRM 70 80 90 100 110 120 PTVQDRLRWM QQIFKYQKNQ IFIHHLVEDG IPSYPNGWQS WSEAVKTLFH EKHFEPSIVF 130 140 150 160 170 180 SSEPQDKAPY EKYLGLEVSL VDPDRTFFNV SATKIRTTPF QYWKFIPKEA RPFFAKTVAI 190 200 210 220 230 240 LGGESSGKSV LVNKLAAVFN TTSAWEYGRE FVFEKLGGDE QAMQYSDYPQ MALGHQRYID 250 260 270 280 290 300 YAVRHSHKIA FIDTDFITTQ AFCIQYEGKA HPFLDSMIKE YPFDVTILLK NNTEWVDDGL 310 320 330 340 350 360 RSLGSQKQRQ QFQQLLKKLL DKYKVPYIEI ESPSYLDRYN QVKAVIEKVL NEEEISELQN 370 TTFPIKGTSQ

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Whole-genome random sequencing and assembly of Haemophilus influenzae Rd." Fleischmann R.D., Adams M.D., White O., Clayton R.A., Kirkness E.F., Kerlavage A.R., Bult C.J., Tomb J.-F., Dougherty B.A., Merrick J.M., McKenney K., Sutton G.G., FitzHugh W., Fields C.A., Gocayne J.D., Scott J.D., Shirley R., Liu L.-I. , Glodek A., Kelley J.M., Weidman J.F., Phillips C.A., Spriggs T., Hedblom E., Cotton M.D., Utterback T.R., Hanna M.C., Nguyen D.T., Saudek D.M., Brandon R.C., Fine L.D., Fritchman J.L., Fuhrmann J.L., Geoghagen N.S.M., Gnehm C.L., McDonald L.A., Small K.V., Fraser C.M., Smith H.O., Venter J.C. Science 269:496-512(1995) [PubMed: 7542800] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: ATCC 51907 / DSM 11121 / KW20 / Rd.
[2]	"Ribosylnicotinamide kinase domain of NadR protein: identification and implications in NAD biosynthesis." Kurnasov O.V., Polanuyer B.M., Ananta S., Sloutsky R., Tam A., Gerdes S.Y., Osterman A.L. J. Bacteriol. 184:6906-6917(2002) [PubMed: 12446641] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, ALTERNATIVE INITIATION, BIOPHYSICOCHEMICAL PROPERTIES. Strain: ATCC 51907 / DSM 11121 / KW20 / Rd.
[3]	"Coupling of NAD+ biosynthesis and nicotinamide ribosyl transport: characterization of NadR ribonucleotide kinase mutants of Haemophilus influenzae." Merdanovic M., Sauer E., Reidl J. J. Bacteriol. 187:4410-4420(2005) [PubMed: 15968050] [Abstract] Cited for: MUTAGENESIS OF LYS-126; GLY-238; TRP-256; TYR-292; ASP-304 AND ARG-352, SUBCELLULAR LOCATION. Strain: ATCC 51907 / DSM 11121 / KW20 / Rd.
[4]	"Crystal structure of Haemophilus influenzae NadR protein. A bifunctional enzyme endowed with NMN adenyltransferase and ribosylnicotinimide kinase activities." Singh S.K., Kurnasov O.V., Chen B., Robinson H., Grishin N.V., Osterman A.L., Zhang H. J. Biol. Chem. 277:33291-33299(2002) [PubMed: 12068016] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.9 ANGSTROMS) OF 57-421 IN COMPLEX WITH NAD, SUBUNIT.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

L42023 Genomic DNA. Translation: AAC22421.1.

PIR

D64091.

RefSeq

NP_438922.1. NC_000907.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1LW7	X-ray	2.90	A	57-421	[»]

ProteinModelPortal

P44308.

SMR

P44308. Positions 57-411.

ModBase

Search...

Protein family/group databases

TCDB

4.B.1.1.2. nicotinamide ribonucleoside (NR) uptake permease (PnuC) family.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

GeneID

950800.

GenomeReviews

Gene locus HI_0763 in contig L42023_GR.

KEGG

hin:HI0763.

NMPDR

fig|71421.1.peg.731.

PATRIC

20190177. VBIHaeInf48452_0802.

TIGR

HI_0763.

Phylogenomic databases

HOGENOM

HBG297196.

OMA

WVDDGLR.

ProtClustDB

PRK08099.

Enzyme and pathway databases

BioCyc

HINF71421:HI_0763-MONOMER.
MetaCyc:MONOMER-8322.

Family and domain databases

InterPro

IPR016429. Bifunc_transcrip_reg_NadR.
IPR004821. Cyt_trans-rel.
IPR006417. NadR_NMN_Atrans.
IPR014729. Rossmann-like_a/b/a_fold.
[Graphical view]

Gene3D

G3DSA:3.40.50.620. Rossmann-like_a/b/a_fold. 1 hit.

KO

K06211.

PIRSF

PIRSF004776. NadR_NMNAT/RNK. 1 hit.

TIGRFAMs

TIGR00125. Cyt_tran_rel. 1 hit.
TIGR01526. NadR_NMN_Atrans. 1 hit.

ProtoNet

Search...

Entry information

Entry name

NADR_HAEIN

Accession

Primary (citable) accession number: P44308

Entry history

Integrated into UniProtKB/Swiss-Prot:	November 1, 1995
Last sequence update:	November 1, 1995
Last modified:	January 25, 2012
This is version 85 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program