2016 Pollard — Actin and Actin-Binding Proteins

Authoritative review: actin polymerization rate constants, profilin/thymosin-β4 monomer pool, Arp2/3 + formin nucleation, cofilin severing, capping. Backbone reference for any STRC actin compute.

Citation

Pollard TD. “Actin and Actin-Binding Proteins.” Cold Spring Harbor Perspectives in Biology, 8: a018226, 2016. doi:10.1101/cshperspect.a018226. PMC4968159. PMID 26988969. Re-ingested from MinerU output 2026-04-25.

TL;DR

Comprehensive review of actin biochemistry and the regulatory protein families (profilin, thymosin-β4, cofilin, Arp2/3, formins, Ena/VASP, capping protein, tropomodulin, gelsolin, cross-linkers, tropomyosin). Establishes the canonical rate constants for monomer-, dimer-, trimer-, and end-specific reactions plus the geometric facts about the actin filament. Single highest-leverage reference for any STRC compute that touches actin assembly, stereocilia treadmilling, or WH2-driven nucleation.

Numbers that matter

Page numbers below refer to the MinerU-converted markdown at ~/BookLibrary/mineru-output/pollard2016/pollard2016/auto/pollard2016.md. Verbatim values from Pollard’s text; secondary citations preserved.

Actin monomer geometry & sequence

ParameterValueSource (PDF page)Notes
Actin polypeptide length375 residuesp.1 (§3)Eukaryotic actin
Subunit half-stagger in filament2.7 nmp.1 (§3)Two-strand right-handed helix
Filament short-pitch helix twist167° per subunitp.1 (§3)Free filament
Subdomain organization4 subdomains, ATP in cleftp.1 (§3)Strong contacts to subdomains 3, 4
Arp ↔ actin sequence identity17–52 %p.1 (§2)Arp1–Arp11

Nucleotide binding (ATP/ADP) on monomeric actin

Reactionk_onk_offK_dSource
ATP binding to nucleotide-free monomer6 µM⁻¹ s⁻¹~10⁻² s⁻¹nanomolar rangep.1 (§4) — De La Cruz & Pollard 1995
Effect of free divalent cation chelation on ATP k_off↑ 20-foldp.1 (§4)

Nucleation kinetics (spontaneous)

StepRate / propertySource
Dimer dissociation rate~10⁶ s⁻¹p.2 (§4) — Cooper 1983; Frieden 1983
Trimer subunit dissociation rate~100 s⁻¹p.2 (§4) — Cooper 1983; Frieden 1983
Stable nucleustetramer (full intermolecular contacts)p.2 (§4)

Filament-end elongation (per Fig. 2 / §4)

EndNucleotidek_on (µM⁻¹ s⁻¹)k_off (s⁻¹)Critical conc.Source
BarbedATP-actin~10 (diffusion-limited)~1~0.1 µMp.2 (§4) — Pollard 1986
PointedATP-actinmuch slower than barbedp.2 (§4)
Both endsMg-ADP-actin1.8 µM (same at both ends)p.2 (§4) — Pollard 1986

ATP hydrolysis on the filament

QuantityValueSource
Mg-ATP hydrolysis rate on monomer7 × 10⁻⁶ s⁻¹p.2 (§4) — Blanchoin & Pollard 2002
Mg-ATP hydrolysis rate on filament subunit0.3 s⁻¹p.2 (§4) — Blanchoin & Pollard 2002
γ-Pi dissociation half-time from polymerized actin~6 minp.2 (§4) — Carlier & Pantaloni 1986
γ-Pi dissociation rate constant~0.003 s⁻¹p.2 (§4) — Carlier & Pantaloni 1986
Pi binding k_on (to ADP-actin filament)~2 M⁻¹ s⁻¹p.2 (§4) — Carlier & Pantaloni 1986
Pi K_d for polymerized ADP-actin~1 mM (pH-dependent)p.2 (§4)

Cellular actin pools

QuantityValueSource
Total actin in cells50–200 µMp.2 (§5)
Unpolymerized fraction~50 % (25–100 µM range)p.2 (§5)
Treadmilling rate at steady state< 1 subunit/secp.2 (§4) — Wegner 1976

Profilin (p.2 §6)

ParameterValueSource
Profilin molecular weight~13–14 kDap.2 (§6)
Profilin K_d for ATP-actin monomer0.1 µMp.2 (§6)
Profilin cellular concentration50–100 µMp.2 (§6)
Profilin K_d at barbed end (ATP-actin filament)> 20 µM (weak)p.2 (§6) — Courtemanche & Pollard 2013
Profilin function on barbed enddoes not block elongation; can slow at high [profilin]p.2 (§6)
Profilin secondary functioncatalyzes ADP→ATP exchange on monomerp.2 (§6) — Mockrin & Korn 1980

Thymosin-β4 (p.2 §6)

ParameterValueSource
Length43 residuesp.2 (§6)
ArchitectureN-terminal helix in barbed-end groove + C-terminal helix at pointed-end cleftp.2 (§6) Fig. 5B
Cellular concentration> 100 µM (leukocytes, platelets)p.2 (§6)
K_d for Mg-ATP-actin~1 µM (micromolar)p.2 (§6)
Functionsequesters monomer; sterically blocks all polymerization-relevant interactionsp.2 (§6)

Cofilin & severing (§7)

ParameterValueSource
Cofilin molecular weight~15 kDap.2 (§7)
Affinity rank for filament subunitsADP > ADP-Pi ≈ ATPp.3 (§7) — Cao 2006
Filament short-pitch twist when cofilin-decorated162° (vs 167° free)p.3 (§7) — McCullough 2011
Long-pitch helix repeat when cofilin-decorated27 nm (vs 36 nm free)p.3 (§7) — McCullough 2011
Severing optimumsub-stoichiometric (interfaces between bare/decorated segments)p.3 (§7) — Andrianantoandro & Pollard 2006; Elam 2013; Ngo 2015
Cofilin Ser3 phosphorylationLIM-kinase ON, Slingshot/chronophin OFF — inactivates all functionsp.3 (§7) — Mizuno 2013

Gelsolin family (§7)

ParameterValueSource
Ca²⁺-binding K_d range0.2 µM (physiological) … >100 µMp.3 (§7) — Nag 2013
Cap locationbarbed end (gelsolin domains 1, 2, 4 long α-helices)p.3 (§7)
PIP₂ effectdissociates gelsolin cap from endp.3 (§7) — Janmey & Stossel 1987

Arp2/3 complex & nucleation (§8)

ParameterValueSource
Subunits7 (Arp2 + Arp3 + 5 novel)p.3 (§8) Fig. 6
Activation requirementbinding to mother filament side + 2 NPF moleculesp.3 (§8) — Padrick 2011; Ti 2011
ExceptionDip1 activates without preexisting filamentp.4 (§8) — Wagner 2013
Inhibitor (drug-like)CK-666p.4 (§8) — Nolen 2009

Formins (§8–9)

ParameterValueSource
Number of mammalian formin isoforms15p.4 (§8) — Higgs & Peterson 2005
Open-state fraction across formins5–90 %p.4 (§9) — Vavylonis 2006
mDia1 open-state fraction90 %p.4 (§9)
mDia1 FH1 polyproline tracks14p.4 (§9)
mDia1 elongation rate (with profilin-actin)700 subunits / sec in fibroblastsp.4 (§9) — Higashida 2004
Speedup vs free barbed end (favorable case)up to 5×p.4 (§9) — Kovar 2006

Ena/VASP (§9)

ParameterValueSource
Oligomeric statetetramerp.4 (§9)
Dwell time on barbed end1.5 secp.4 (§9) — much less processive than formins
Functionpromotes elongation, inhibits capping; no detectable nucleationp.4 (§9)

Capping protein (§10)

ParameterValueSource
Architectureα/β heterodimerp.4 (§10)
Cellular concentrationlow micromolarp.4 (§10)
Half-time for dissociation from barbed end30 minp.4 (§10)
RegulatorsCARMIL (allosteric); polyphosphoinositides (steric); V-1/myotrophin (sequester)p.4 (§10) — Edwards 2014

Tropomodulin (§10)

ParameterValueSource
Specificityexclusive pointed-end capp.4 (§10) — Rao 2014
Architecturewraps the three terminal pointed-end subunitsp.4 (§10)
Stabilizationbinds N-termini of two tropomyosin molecules → strengthens capp.4 (§10)
Functionblocks both addition AND loss; allows slow exchange in vivop.4 (§10)

Cross-linker ABDs (§11)

ParameterValueSource
Typical ABD K_d for F-actin~10 µMp.5 (§11)
Exchange timescalesub-secondp.5 (§11)
Mechanical consequencenetworks stiff under fast deformation, deformable on tens-of-seconds timescalep.5 (§11) — Yao 2011; Xu 1998

Why this matters for STRC

  1. h09 hydrogel — every parameter the WH2-bundling models use about actin (free monomer concentration, profilin/Tβ4 partitioning, treadmilling rate, ABD-class F-actin K_d as proxy) is in this one review. The note [[Recipe — Profilin Thymosin-β4 Monomer Pool Partitioning]] extracts the partitioning algorithm from §6.
  2. Stereocilia mechanics — tropomodulin at the pointed end of stereocilia thin filaments is exactly the §10 tropomodulin mechanism. Capping protein 30-min off-rate (§10) is the right timescale to compare against bundle turnover.
  3. Treadmilling — pointed-end vs barbed-end critical-concentration asymmetry (§4 + §10 Fig. 8) explains why stereocilia rootlets at the base disassemble while tips grow.
  4. Cofilin filament geometry change (167°→162°, 36→27 nm) is not just a chemistry detail — it’s a mechanical-state change that any AFM/FEM model of stereocilia under cofilin regulation must account for. See [[Cofilin Filament Twist Geometry Change]].
  5. Arp2/3 needs TWO NPFs — relevant if any STRC delivery strategy invokes Arp2/3-driven branched-network seeding. Simple “one WASp” models are wrong.

Atomic notes derived from this paper

  • [[Actin Polymerization Kinetics Reference Table]] — P0 reference (consolidated from §4 + §6 + §10)
  • [[Cofilin Filament Twist Geometry Change]] — P0 (§7 mechanical state change)
  • [[Capping Protein Dissociation Half-Life]] — P0 (§10, relevance to stereocilia caps)
  • [[Recipe — Profilin Thymosin-β4 Monomer Pool Partitioning]] — P1
  • [[Tropomodulin Pointed End Cap Mechanism]] — P2 (§10)
  • [[Cofilin Severing Substoichiometric Optimum]] — P2 (§7)
  • [[Arp2 3 Complex Two-NPF Activation Requirement]] — P2 (§8)

Relevance to hydrogel model (legacy section, retained)

The “K_d ~10 µM for ABD-class cross-linkers” (§11) is the best available proxy for WH2 × F-actin side-binding in the absence of direct measurement. The model’s WH2_KD_FACTIN_M = 5 µM is therefore in the right ballpark as an optimistic estimate but has no primary measurement backing. A more conservative estimate would be 10–100 µM or higher. WH2 itself (§6) binds in the barbed-end groove of monomeric actin — the side-binding affinity is an extrapolation, not a direct Pollard 2016 measurement.

Connections