Influence of nucleotide binding site occupancy on the thermal stability of the F1 portion of the chloroplast ATP synthase.


Abstract

The irreversible thermal denaturation of the F1 portion of the chloroplast ATP synthase (CF1) was examined by differential scanning calorimetry, ATPase activity loss, and release of bound nucleotides. In nearly all cases, the loss of ATPase activity closely paralleled the temperature dependence of the excess heat capacity. Although the irreversible nature of the denaturation precluded thermodynamic interpretation, a kinetic analysis was feasible. A two-state kinetic model was found to fit the calorimetric data very well. The activation energies of thermal denaturation calculated from calorimetric data were very close to those determined from Arrhenius plots of the apparent first-order rate constants of loss of ATPase activity versus reciprocal of the temperature. The nucleotide binding site occupancy profoundly influenced the temperature at which thermal denaturation occurred. In particular, the temperature at which the maximum in excess heat capacity occurs (Tm) was increased about 8 degrees C by occupancy of tight, noncatalytic ATP binding sites and by an additional 3-4 degrees C by the presence of nucleotides in the medium during heating. The thermal denaturation of CF1, an enzyme composed of nine polypeptide chains, is highly cooperative in that it obeys the simple two-step kinetic model. Since the removal of the epsilon and delta subunits has little effect on thermal denaturation, the major forces that stabilize CF1 must, thus, be between the alpha, beta, and gamma subunits. Study holds ProTherm entries: 5225 Extra Details:

Submission Details

ID: 6md7C7WG4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:29 p.m.

Version: 1

Publication Details
Wang ZY;Freire E;McCarty RE,J. Biol. Chem. (1993) Influence of nucleotide binding site occupancy on the thermal stability of the F1 portion of the chloroplast ATP synthase. PMID:8407905
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
6FKF 2018-05-23 3.15 Chloroplast F1Fo conformation 1
1FX0 2001-09-25 3.2 Crystal structure of the chloroplast F1-ATPase from spinach
1KMH 2002-03-13 3.4 Crystal Structure of spinach chloroplast F1-ATPase complexed with tentoxin
6FKH 2018-05-23 4.2 Chloroplast F1Fo conformation 2
6FKI 2018-05-23 4.3 Chloroplast F1Fo conformation 3

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.1 ATP synthase subunit alpha, chloroplastic A7M951 ATPA_CUSRE
90.1 ATP synthase subunit alpha, chloroplastic A8W3A9 ATPA_CUSEX
90.8 ATP synthase subunit alpha, chloroplastic P08215 ATPA_PEA
91.1 ATP synthase subunit alpha, chloroplastic Q06RE6 ATPA_JASNU
91.9 ATP synthase subunit alpha, chloroplastic B0Z4W6 ATPA_OENBI
91.3 ATP synthase subunit alpha, chloroplastic A7Y3A4 ATPA_IPOPU
91.7 ATP synthase subunit alpha, chloroplastic A6MMJ2 ATPA_DIOEL
91.7 ATP synthase subunit alpha, chloroplastic B0Z550 ATPA_OENGL
91.9 ATP synthase subunit alpha, chloroplastic Q9MTL7 ATPA_OENEH
91.3 ATP synthase subunit alpha, chloroplastic Q3BAQ7 ATPA_PHAAO
92.1 ATP synthase subunit alpha, chloroplastic B0Z5D4 ATPA_OENPA
92.1 ATP synthase subunit alpha, chloroplastic B0Z4N2 ATPA_OENAR
90.9 ATP synthase subunit alpha, chloroplastic A9L981 ATPA_LEMMI
91.5 ATP synthase subunit alpha, chloroplastic Q06GS5 ATPA_PIPCE
92.1 ATP synthase subunit alpha, chloroplastic Q70XV0 ATPA_AMBTC
92.3 ATP synthase subunit alpha, chloroplastic Q0G9N4 ATPA_LIRTU
91.9 ATP synthase subunit alpha, chloroplastic Q06H12 ATPA_DRIGR
92.7 ATP synthase subunit alpha, chloroplastic A8SE59 ATPA_CERDE
92.5 ATP synthase subunit alpha, chloroplastic A6MMS9 ATPA_ILLOL
92.9 ATP synthase subunit alpha, chloroplastic A4QLR8 ATPA_NASOF
92.5 ATP synthase subunit alpha, chloroplastic A4QL04 ATPA_DRANE
92.3 ATP synthase subunit alpha, chloroplastic A4QJR8 ATPA_OLIPU
92.5 ATP synthase subunit alpha, chloroplastic Q6EW63 ATPA_NYMAL
92.7 ATP synthase subunit alpha, chloroplastic A1XFU0 ATPA_NUPAD
92.5 ATP synthase subunit alpha, chloroplastic A4QKR6 ATPA_CRUWA
92.3 ATP synthase subunit alpha, chloroplastic A4QKH7 ATPA_CAPBU
93.1 ATP synthase subunit alpha, chloroplastic Q3V549 ATPA_ACOCL
93.1 ATP synthase subunit alpha, chloroplastic A9LYH0 ATPA_ACOAM
92.7 ATP synthase subunit alpha, chloroplastic A4QK90 ATPA_BARVE
92.9 ATP synthase subunit alpha, chloroplastic Q7YJY4 ATPA_CALFG
92.5 ATP synthase subunit alpha, chloroplastic P56757 ATPA_ARATH
92.9 ATP synthase subunit alpha, chloroplastic Q09FX6 ATPA_NANDO
92.7 ATP synthase subunit alpha, chloroplastic A4QLH9 ATPA_LOBMA
93.3 ATP synthase subunit alpha, chloroplastic A4GGB2 ATPA_PHAVU
92.9 ATP synthase subunit alpha, chloroplastic Q2QDA3 ATPA_CUCSA
93.3 ATP synthase subunit alpha, chloroplastic A6MMA7 ATPA_CHLSC
92.9 ATP synthase subunit alpha, chloroplastic A4QK03 ATPA_ARAHI
92.5 ATP synthase subunit alpha, chloroplastic A4QJA0 ATPA_AETCO
93.1 ATP synthase subunit alpha, chloroplastic A4QL91 ATPA_LEPVR
92.7 ATP synthase subunit alpha, chloroplastic A4QJI4 ATPA_AETGR
93.7 ATP synthase subunit alpha, chloroplastic Q9BBS3 ATPA_LOTJA
93.9 ATP synthase subunit alpha, chloroplastic B1A920 ATPA_CARPA
93.9 ATP synthase subunit alpha, chloroplastic A4GYP3 ATPA_POPTR
93.9 ATP synthase subunit alpha, chloroplastic Q14FH2 ATPA_POPAL
93.9 ATP synthase subunit alpha, chloroplastic Q2PMS8 ATPA_SOYBN
93.7 ATP synthase subunit alpha, chloroplastic Q09MJ3 ATPA_CITSI
93.9 ATP synthase subunit alpha, chloroplastic Q09X32 ATPA_MORIN
93.5 ATP synthase subunit alpha, chloroplastic A1XGM3 ATPA_RANMC
94.1 ATP synthase subunit alpha, chloroplastic A0ZZ20 ATPA_GOSBA
94.3 ATP synthase subunit alpha, chloroplastic A6MM21 ATPA_BUXMI
94.1 ATP synthase subunit alpha, chloroplastic Q332Y4 ATPA_LACSA
93.7 ATP synthase subunit alpha, chloroplastic Q1KXW5 ATPA_HELAN
94.1 ATP synthase subunit alpha, chloroplastic Q68S21 ATPA_PANGI
94.5 ATP synthase subunit alpha, chloroplastic A0A320 ATPA_COFAR
94.5 ATP synthase subunit alpha, chloroplastic Q2L8Z1 ATPA_GOSHI
94.3 ATP synthase subunit alpha, chloroplastic Q0ZJ35 ATPA_VITVI
94.7 ATP synthase subunit alpha, chloroplastic Q33C53 ATPA_NICTO
94.7 ATP synthase subunit alpha, chloroplastic P00823 ATPA_TOBAC
94.5 ATP synthase subunit alpha, chloroplastic Q27S65 ATPA_SOLTU
94.5 ATP synthase subunit alpha, chloroplastic Q2MIB5 ATPA_SOLLC
94.5 ATP synthase subunit alpha, chloroplastic Q2MIK2 ATPA_SOLBU
94.7 ATP synthase subunit alpha, chloroplastic Q09G61 ATPA_PLAOC
94.7 ATP synthase subunit alpha, chloroplastic Q3C1H4 ATPA_NICSY
94.3 ATP synthase subunit alpha, chloroplastic Q0G9X7 ATPA_DAUCA
94.5 ATP synthase subunit alpha, chloroplastic Q8S8Y3 ATPA_ATRBE
94.9 ATP synthase subunit alpha, chloroplastic B1NWD5 ATPA_MANES
95.1 ATP synthase subunit alpha, chloroplastic Q589B3 ATPA_SILLA
95.5 ATP synthase subunit alpha, chloroplastic Q49L13 ATPA_EUCGG
100.0 ATP synthase subunit alpha, chloroplastic P06450 ATPA_SPIOL